“Dr. Julie Eggington and I worked together in developing the variant science approach to categorizing variants in vast datasets obtained from next generation sequencing at Courtagen Life Sciences. Reporting variant classifications for large datasets is at the forefront of current genetic diagnostic testing and Julie’s extensive and deep background in every aspect of variant classification was extremely valuable to creating a process of the highest standards. I have yet to find a problem to which Julie wasn’t able to find the solution and with deep expertise, eloquence and grace to boot. She is a master manager with a thoughtful entrepreneurial spirit and ethical to the core. Julie is also a dynamic speaker with the unique ability to present complex material to any audience. It was my distinct pleasure to be her manager and I sincerely hope I have the opportunity to work with Julie again. ”
About
The NIH warns that “Most genetic tests today are not regulated, meaning that they go to…
Activity
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Our first curated draft somatic structural variant benchmark for the new GIAB PDAC tumor cell line HG008-T is at https://lnkd.in/exMpnm3m, based on…
Our first curated draft somatic structural variant benchmark for the new GIAB PDAC tumor cell line HG008-T is at https://lnkd.in/exMpnm3m, based on…
Liked by Julie Eggington, MS, PhD
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I was very honored to receive 2024 Anna and Edward Nevius Award from the FDA Statistical Association (FDASA) last Friday. This award was created in…
I was very honored to receive 2024 Anna and Edward Nevius Award from the FDA Statistical Association (FDASA) last Friday. This award was created in…
Liked by Julie Eggington, MS, PhD
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I am so thankful for the opportunity that I had to attend the University of Florida International Aortic Symposium and present my center’s poster on,…
I am so thankful for the opportunity that I had to attend the University of Florida International Aortic Symposium and present my center’s poster on,…
Liked by Julie Eggington, MS, PhD
Experience
Education
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University of Wisconsin-Madison
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Julie received her Ph.D. in the laboratory of Dr. Michael Cox researching the molecular biology of DNA homologous recombination. Her particular focus was investigating this pathway in Deinococcus radiodurans, focusing on the SSB protein. Julie received a fellowship and other academic awards during her studies.
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Julie received her Masters in the laboratory of Dr. Bradley Preston. Her research focused on DNA mismatch repair, polymerase proofreading, and nucleotide pool maintenance and their roles in maintaining genomic stability to avoid tumorigenesis.
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Activities and Societies: Golden Key International Honour Society
During Julie's undergraduate studies she was able to work in the laboratories of Dr. Michael Whiting (phylogenetics), Dr. Steven Fleming (organic chemistry), and Dr. Gary Frederick (organic chemistry, Hawaii campus). Julie received multiple awards and scholarships during her studies.
Publications
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The Science and Art of Clinical Genetic Variant Classification and Its Impact on Test Accuracy
Annual Review of Genomics and Human Genetics
Hunter H. Giles, Madhuri R. Hegde, Elaine Lyon, Christine M. Stanley, Iain D. Kerr, Megan E. Garlapow, Julie M. Eggington
Abstract
Clinical genetic variant classification science is a growing subspecialty of clinical genetics and genomics. The field's continued improvement is essential for the success of precision medicine in both germline (hereditary) and somatic (oncology) contexts. This review focuses on variant classification for DNA next-generation sequencing tests. We first…Hunter H. Giles, Madhuri R. Hegde, Elaine Lyon, Christine M. Stanley, Iain D. Kerr, Megan E. Garlapow, Julie M. Eggington
Abstract
Clinical genetic variant classification science is a growing subspecialty of clinical genetics and genomics. The field's continued improvement is essential for the success of precision medicine in both germline (hereditary) and somatic (oncology) contexts. This review focuses on variant classification for DNA next-generation sequencing tests. We first summarize current limitations in variant discovery and definition, and then describe the current five- and four-tier classification systems outlined in dominant standards and guideline publications for germline and somatic tests, respectively. We then discuss measures of variant classification discordance and the field's bias for positive results, as well as considerations for panel size and population screening in the context of estimates of positive predictive value that incorporate estimated variant classification imperfections. Finally, we share opinions on the current state of variant classification from some of the authors of the most widely used standards and guideline publications and from other domain experts. Expected final online publication date for the Annual Review of Genomics and Human Genetics Volume 22 is August 2021. -
Assessment of in silico protein sequence analysis in the clinical classification of variants in cancer risk genes
Journal of Community Genetics. Kerr ID, Cox HC, Moyes K, Evans B, Burdett BC, van Kan A, McElroy H, Vail PJ, Brown KL, Sumampong DB, Monteferrante NJ, Hardman KL, Theisen A, Mundt E, Wenstrup RJ, Eggington JM.
Abstract
Missense variants represent a significant proportion of variants identified in clinical genetic testing. In the absence of strong clinical or functional evidence, the American College of Medical Genetics recommends that these findings be classified as variants of uncertain significance (VUS). VUSs may be reclassified to better inform patient care when new evidence is available. It is critical that the methods used for reclassification are robust in order to prevent inappropriate…Abstract
Missense variants represent a significant proportion of variants identified in clinical genetic testing. In the absence of strong clinical or functional evidence, the American College of Medical Genetics recommends that these findings be classified as variants of uncertain significance (VUS). VUSs may be reclassified to better inform patient care when new evidence is available. It is critical that the methods used for reclassification are robust in order to prevent inappropriate medical management strategies and unnecessary, life-altering surgeries. In an effort to provide evidence for classification, several in silico algorithms have been developed that attempt to predict the functional impact of missense variants through amino acid sequence conservation analysis. We report an analysis comparing internally derived, evidence-based classifications with the results obtained from six commonly used algorithms. We compiled a dataset of 1118 variants in BRCA1, BRCA2, MLH1, and MSH2 previously classified by our laboratory's evidence-based variant classification program. We compared internally derived classifications with those obtained from the following in silico tools: Align-GVGD, CONDEL, Grantham Analysis, MAPP-MMR, PolyPhen-2, and SIFT. Despite being based on similar underlying principles, all algorithms displayed marked divergence in accuracy, specificity, and sensitivity. Overall, accuracy ranged from 58.7 to 90.8% while the Matthews Correlation Coefficient ranged from 0.26-0.65. CONDEL, a weighted average of multiple algorithms, did not perform significantly better than its individual components evaluated here. These results suggest that the in silico algorithms evaluated here do not provide reliable evidence regarding the clinical significance of missense variants in genes associated with hereditary cancer. -
Hurt, tired and queasy: Specific variants in the ATPase domain of the TRAP1 mitochondrial chaperone are associated with common, chronic "functional" symptomatology including pain, fatigue and gastrointestinal dysmotility
Mitochondrion. Boles RG, Hornung HA, Moody AE, Ortiz TB, Wong SA, Eggington JM, Stanley CM, Gao M, Zhou H, McLaughlin S, Zare AS, Sheldon KM, Skolnick J, McKernan KJ.
Abstract
Functional disorders are common conditions with a substantial impact on a patients' wellbeing, and can be diagnostically elusive. There are bidirectional associations between functional disorders and mitochondrial dysfunction. In this study, provided clinical information and the exon sequence of the TRAP1 mitochondrial chaperone were retrospectively reviewed with a focus on the functional categories of chronic pain, fatigue and gastrointestinal dysmotility. Very-highly conserved…Abstract
Functional disorders are common conditions with a substantial impact on a patients' wellbeing, and can be diagnostically elusive. There are bidirectional associations between functional disorders and mitochondrial dysfunction. In this study, provided clinical information and the exon sequence of the TRAP1 mitochondrial chaperone were retrospectively reviewed with a focus on the functional categories of chronic pain, fatigue and gastrointestinal dysmotility. Very-highly conserved TRAP1 variants were identified in 73 of 930 unrelated patients. Functional symptomatology is strongly associated with specific variants in the ATPase binding pocket. In particular, the combined presence of all three functional categories is strongly associated with p.Ile253Val (OR 7.5, P=0.0001) and with two other interacting variants (OR 18, P=0.0005). Considering a 1-2% combined variant prevalence and high odds ratios, these variants may be an important factor in the etiology of functional symptomatology. -
Comparison of locus-specific databases for BRCA1 and BRCA2 variants reveals disparity in variant classification within and among databases
Journal of Community Genetics. Vail PJ, Morris B, van Kan A, Burdett BC, Moyes K, Theisen A, Kerr ID, Wenstrup RJ, Eggington JM.
Abstract
Genetic variants of uncertain clinical significance (VUSs) are a common outcome of clinical genetic testing. Locus-specific variant databases (LSDBs) have been established for numerous disease-associated genes as a research tool for the interpretation of genetic sequence variants to facilitate variant interpretation via aggregated data. If LSDBs are to be used for clinical practice, consistent and transparent criteria regarding the deposition and interpretation of variants are…Abstract
Genetic variants of uncertain clinical significance (VUSs) are a common outcome of clinical genetic testing. Locus-specific variant databases (LSDBs) have been established for numerous disease-associated genes as a research tool for the interpretation of genetic sequence variants to facilitate variant interpretation via aggregated data. If LSDBs are to be used for clinical practice, consistent and transparent criteria regarding the deposition and interpretation of variants are vital, as variant classifications are often used to make important and irreversible clinical decisions. In this study, we performed a retrospective analysis of 2017 consecutive BRCA1 and BRCA2 genetic variants identified from 24,650 consecutive patient samples referred to our laboratory to establish an unbiased dataset representative of the types of variants seen in the US patient population, submitted by clinicians and researchers for BRCA1 and BRCA2 testing. We compared the clinical classifications of these variants among five publicly accessible BRCA1 and BRCA2 variant databases: BIC, ClinVar, HGMD (paid version), LOVD, and the UMD databases. Our results show substantial disparity of variant classifications among publicly accessible databases. Furthermore, it appears that discrepant classifications are not the result of a single outlier but widespread disagreement among databases. This study also shows that databases sometimes favor a clinical classification when current best practice guidelines (ACMG/AMP/CAP) would suggest an uncertain classification. Although LSDBs have been well established for research applications, our results suggest several challenges preclude their wider use in clinical practice. -
Development and validation of a new algorithm for the reclassification of genetic variants identified in the BRCA1 and BRCA2 genes
Breast Cancer Research and Treatment. Pruss D, Morris B, Hughes E, Eggington JM, Esterling L, Robinson BS, van Kan A, Fernandes PH, Roa BB, Gutin A, Wenstrup RJ, Bowles KR.
Abstract
BRCA1 and BRCA2 sequencing analysis detects variants of uncertain clinical significance in approximately 2 % of patients undergoing clinical diagnostic testing in our laboratory. The reclassification of these variants into either a pathogenic or benign clinical interpretation is critical for improved patient management. We developed a statistical variant reclassification tool based on the premise that probands with disease-causing mutations are expected to have more severe personal…Abstract
BRCA1 and BRCA2 sequencing analysis detects variants of uncertain clinical significance in approximately 2 % of patients undergoing clinical diagnostic testing in our laboratory. The reclassification of these variants into either a pathogenic or benign clinical interpretation is critical for improved patient management. We developed a statistical variant reclassification tool based on the premise that probands with disease-causing mutations are expected to have more severe personal and family histories than those having benign variants. The algorithm was validated using simulated variants based on approximately 145,000 probands, as well as 286 BRCA1 and 303 BRCA2 true variants. Positive and negative predictive values of ≥99 % were obtained for each gene. Although the history weighting algorithm was not designed to detect alleles of lower penetrance, analysis of the hypomorphic mutations c.5096G>A (p.Arg1699Gln; BRCA1) and c.7878G>C (p.Trp2626Cys; BRCA2) indicated that the history weighting algorithm is able to identify some lower penetrance alleles. The history weighting algorithm is a powerful tool that accurately assigns actionable clinical classifications to variants of uncertain clinical significance. While being developed for reclassification of BRCA1 and BRCA2 variants, the history weighting algorithm is expected to be applicable to other cancer- and non-cancer-related genes. -
A comprehensive laboratory-based program for classification of variants of uncertain significance in hereditary cancer genes.
Clinical Genetics. 2013. doi: 10.1111/cge.12315. Eggington JM, Bowles KR, Moyes K, Manley S, Esterling L, Sizemore S, Rosenthal E, Theisen A, Saam J, Arnell C, Pruss D, Bennett J, Burbidge LA, Roa B, Wenstrup RJ.
Abstract
Genetic testing has the potential to guide the prevention and treatment of disease in a variety of settings, and recent technical advances have greatly increased our ability to acquire large amounts of genetic data. The interpretation of this data remains challenging, as the clinical significance of genetic variation detected in the laboratory is not always clear. Although regulatory agencies and professional societies provide some guidance regarding the classification, reporting…Abstract
Genetic testing has the potential to guide the prevention and treatment of disease in a variety of settings, and recent technical advances have greatly increased our ability to acquire large amounts of genetic data. The interpretation of this data remains challenging, as the clinical significance of genetic variation detected in the laboratory is not always clear. Although regulatory agencies and professional societies provide some guidance regarding the classification, reporting, and long-term follow-up of variants, few protocols for the implementation of these guidelines have been described. Because the primary aim of clinical testing is to provide results to inform medical management, a variant classification program that offers timely, accurate, confident and cost-effective interpretation of variants should be an integral component of the laboratory process. Here we describe the components of our laboratory's current variant classification program (VCP), based on 20 years of experience and over one million samples tested, using the BRCA1/2 genes as a model. Our VCP has lowered the percentage of tests in which one or more BRCA1/2 variants of uncertain significance (VUSs) are detected to 2.1% in the absence of a pathogenic mutation, demonstrating how the coordinated application of resources toward classification and reclassification significantly impacts the clinical utility of testing. -
Functional evaluation of BRCA2 variants mapping to the PALB2-binding and C-terminal DNA-binding domains using a mouse ES cell-based assay
Human Molecular Genetics. 2012. 21(18):3993-4006. Biswas K, Das R, Eggington JM, Qiao H, North SL, Stauffer S, Burkett SS, Martin BK, Southon E, Sizemore SC, Pruss D, Bowles KR, Roa BB, Hunter N, Tessarollo L, Wenstrup RJ, Byrd RA, Sharan SK.
Abstract
Single-nucleotide substitutions and small in-frame insertions or deletions identified in human breast cancer susceptibility genes BRCA1 and BRCA2 are frequently classified as variants of unknown clinical significance (VUS) due to the availability of very limited information about their functional consequences. Such variants can most reliably be classified as pathogenic or non-pathogenic based on the data of their co-segregation with breast cancer in affected families and/or their…Abstract
Single-nucleotide substitutions and small in-frame insertions or deletions identified in human breast cancer susceptibility genes BRCA1 and BRCA2 are frequently classified as variants of unknown clinical significance (VUS) due to the availability of very limited information about their functional consequences. Such variants can most reliably be classified as pathogenic or non-pathogenic based on the data of their co-segregation with breast cancer in affected families and/or their co-occurrence with a pathogenic mutation. Biological assays that examine the effect of variants on protein function can provide important information that can be used in conjunction with available familial data to determine the pathogenicity of VUS. In this report, we have used a previously described mouse embryonic stem (mES) cell-based functional assay to characterize eight BRCA2 VUS that affect highly conserved amino acid residues and map to the N-terminal PALB2-binding or the C-terminal DNA-binding domains. For several of these variants, very limited co-segregation information is available, making it difficult to determine their pathogenicity. Based on their ability to rescue the lethality of Brca2-deficient mES cells and their effect on sensitivity to DNA-damaging agents, homologous recombination and genomic integrity, we have classified these variants as pathogenic or non-pathogenic. In addition, we have used homology-based modeling as a predictive tool to assess the effect of some of these variants on the structural integrity of the C-terminal DNA-binding domain and also generated a knock-in mouse model to analyze the physiological significance of a residue reported to be essential for the interaction of BRCA2 with meiosis-specific recombinase, DMC1. -
Thirty-nine-year-old with familial colon cancer, and variant of undetermined significance in MSH6
Seminars in Oncology. 2012. 39(2):125-131. Adonizio C, Gazzillo M, Knezetic J, Snyder C, Lynch HT, Rybak C, Hall MJ, Lowstuter K, Eggington J, Morris GJ.
A case study from different perspectives.
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Mutator Suppression and Escape from Replication Error-Induced Extinction in Yeast
PLoS Genetics. 2011. 7(10):e1002282. Herr AJ, Ogawa M, Lawrence NA, Williams LN, Eggington JM, Singh M, Smith RA, Preston BD.
Abstract
Cells rely on a network of conserved pathways to govern DNA replication fidelity. Loss of polymerase proofreading or mismatch repair elevates spontaneous mutation and facilitates cellular adaptation. However, double mutants are inviable, suggesting that extreme mutation rates exceed an error threshold. Here we combine alleles that affect DNA polymerase δ (Pol δ) proofreading and mismatch repair to define the maximal error rate in haploid yeast and to characterize genetic suppressors…Abstract
Cells rely on a network of conserved pathways to govern DNA replication fidelity. Loss of polymerase proofreading or mismatch repair elevates spontaneous mutation and facilitates cellular adaptation. However, double mutants are inviable, suggesting that extreme mutation rates exceed an error threshold. Here we combine alleles that affect DNA polymerase δ (Pol δ) proofreading and mismatch repair to define the maximal error rate in haploid yeast and to characterize genetic suppressors of mutator phenotypes. We show that populations tolerate mutation rates 1,000-fold above wild-type levels but collapse when the rate exceeds 10⁻³ inactivating mutations per gene per cell division. Variants that escape this error-induced extinction (eex) rapidly emerge from mutator clones. One-third of the escape mutants result from second-site changes in Pol δ that suppress the proofreading-deficient phenotype, while two-thirds are extragenic. The structural locations of the Pol δ changes suggest multiple antimutator mechanisms. Our studies reveal the transient nature of eukaryotic mutators and show that mutator phenotypes are readily suppressed by genetic adaptation. This has implications for the role of mutator phenotypes in cancer. -
Predicting sites of ADAR editing in double-stranded RNA
Nature Communications. 2011. 2:319. Eggington JM, Greene T, Bass BL.
Abstract
ADAR (adenosine deaminase that acts on RNA) editing enzymes target coding and noncoding double-stranded RNA (dsRNA) and are essential for neuronal function. Early studies showed that ADARs preferentially target adenosines with certain 5' and 3' neighbours. Here we use current Sanger sequencing protocols to perform a more accurate and quantitative analysis. We quantified editing sites in an ∼800-bp dsRNA after reaction with human ADAR1 or ADAR2, or their catalytic domains alone…Abstract
ADAR (adenosine deaminase that acts on RNA) editing enzymes target coding and noncoding double-stranded RNA (dsRNA) and are essential for neuronal function. Early studies showed that ADARs preferentially target adenosines with certain 5' and 3' neighbours. Here we use current Sanger sequencing protocols to perform a more accurate and quantitative analysis. We quantified editing sites in an ∼800-bp dsRNA after reaction with human ADAR1 or ADAR2, or their catalytic domains alone. These large data sets revealed that neighbour preferences are mostly dictated by the catalytic domain, but ADAR2's dsRNA-binding motifs contribute to 3' neighbour preferences. For all proteins, the 5' nearest neighbour was most influential, but adjacent bases also affected editing site choice. We developed algorithms to predict editing sites in dsRNA of any sequence, and provide a web-based application. The predictive power of the algorithm on fully base-paired dsRNA, compared with biological substrates containing mismatches, bulges and loops, elucidates structural contributions to editing specificity. -
Binding of the Dimeric Deinococcus radiodurans Single-stranded DNA Binding Protein to Single-stranded DNA
Biochemistry. 2010. 49(38):8266-8275. Kozlov AG, Eggington JM, Cox MM, Lohman TM.
Abstract
Deinococcus radiodurans single-stranded (ss) DNA binding protein (DrSSB) originates from a radiation-resistant bacterium and participates in DNA recombination, replication, and repair. Although it functions as a homodimer, it contains four DNA binding domains (OB-folds) and thus is structurally similar to the Escherichia coli SSB (EcoSSB) homotetramer. We examined the equilibrium binding of DrSSB to ssDNA for comparison with that of EcoSSB. We find that the occluded site size of…Abstract
Deinococcus radiodurans single-stranded (ss) DNA binding protein (DrSSB) originates from a radiation-resistant bacterium and participates in DNA recombination, replication, and repair. Although it functions as a homodimer, it contains four DNA binding domains (OB-folds) and thus is structurally similar to the Escherichia coli SSB (EcoSSB) homotetramer. We examined the equilibrium binding of DrSSB to ssDNA for comparison with that of EcoSSB. We find that the occluded site size of DrSSB on poly(dT) is ∼45 nucleotides under low-salt conditions (<0.02 M NaCl) but increases to 50-55 nucleotides at ≥0.2 M NaCl. This suggests that DrSSB undergoes a transition between ssDNA binding modes, which is observed for EcoSSB, although the site size difference between modes is not as large as for EcoSSB, suggesting that the pathways of ssDNA wrapping differ for these two proteins. The occluded site size corresponds well to the contact site size (52 nucleotides) determined by isothermal titration calorimetry (ITC). Electrophoretic studies of complexes of DrSSB with phage M13 ssDNA indicate the formation of stable, highly cooperative complexes under low-salt conditions. Using ITC, we find that DrSSB binding to oligo(dT)s with lengths close to the determined site size (50-55 nucleotides) is stoichiometric with a ΔH(obs) of approximately -94 ± 4 kcal/mol, somewhat smaller than that for EcoSSB (approximately -130 kcal/mol) under the same conditions. The observed binding enthalpy shows a large sensitivity to NaCl concentration, similar to that observed for EcoSSB. With the exception of the less dramatic change in occluded site size, the behavior of DrSSB is similar to that of EcoSSB protein (although clear quantitative differences exist). These common features for SSB proteins having multiple DNA binding domains enable versatility of SSB function in vivo. -
Directed Evolution of Ionizing Radiation Resistance in Escherichia coli
Journal of Bacteriology. 2009. 191(16):5240-5252. Harris, Pollock, Wood, Goiffon, Klingele, Cabo, Schackwitz, Martin, Eggington J, Durfee, Middle, Norton, Popelars, Li, Klugman, Hamilton, Bane, Pennacchio, Albert, Perna, Cox MM, Battista JR.
Abstract
We have generated extreme ionizing radiation resistance in a relatively sensitive bacterial species, Escherichia coli, by directed evolution. Four populations of Escherichia coli K-12 were derived independently from strain MG1655, with each specifically adapted to survive exposure to high doses of ionizing radiation. D(37) values for strains isolated from two of the populations approached that exhibited by Deinococcus radiodurans. Complete genomic sequencing was carried out on nine…Abstract
We have generated extreme ionizing radiation resistance in a relatively sensitive bacterial species, Escherichia coli, by directed evolution. Four populations of Escherichia coli K-12 were derived independently from strain MG1655, with each specifically adapted to survive exposure to high doses of ionizing radiation. D(37) values for strains isolated from two of the populations approached that exhibited by Deinococcus radiodurans. Complete genomic sequencing was carried out on nine purified strains derived from these populations. Clear mutational patterns were observed that both pointed to key underlying mechanisms and guided further characterization of the strains. In these evolved populations, passive genomic protection is not in evidence. Instead, enhanced recombinational DNA repair makes a prominent but probably not exclusive contribution to genome reconstitution. Multiple genes, multiple alleles of some genes, multiple mechanisms, and multiple evolutionary pathways all play a role in the evolutionary acquisition of extreme radiation resistance. Several mutations in the recA gene and a deletion of the e14 prophage both demonstrably contribute to and partially explain the new phenotype. Mutations in additional components of the bacterial recombinational repair system and the replication restart primosome are also prominent, as are mutations in genes involved in cell division, protein turnover, and glutamate transport. At least some evolutionary pathways to extreme radiation resistance are constrained by the temporally ordered appearance of specific alleles. -
Polar destabilization of DNA duplexes with single-stranded overhangs by the Deinococcus radiodurans SSB protein
Biochemistry. 2006. 45:14490-14502. Eggington JM, Kozlov AG, Cox MM, Lohman TM.
Abstract
The Deinococcus radiodurans SSB protein has an occluded site size of 50 +/- 2 nucleotides on ssDNA but can form a stable complex with a 26-30-nucleotide oligodeoxynucleotide using a subset of its four ssDNA binding domains. Quantitative estimates of D. radiodurans SSB protein in the D. radiodurans cell indicate approximately 2500-3000 dimers/cell, independent of the level of irradiation. At biologically relevant concentrations, when bound at single-strand-double-strand DNA junctions…Abstract
The Deinococcus radiodurans SSB protein has an occluded site size of 50 +/- 2 nucleotides on ssDNA but can form a stable complex with a 26-30-nucleotide oligodeoxynucleotide using a subset of its four ssDNA binding domains. Quantitative estimates of D. radiodurans SSB protein in the D. radiodurans cell indicate approximately 2500-3000 dimers/cell, independent of the level of irradiation. At biologically relevant concentrations, when bound at single-strand-double-strand DNA junctions in vitro, D. radiodurans SSB protein has a limited capacity to displace the shorter strand of the duplex, permitting it to bind to single-strand extensions shorter than 26-30 nucleotides. The capacity to displace the shorter strand of the duplex shows a pronounced bias for extensions with a free 3' end. The Escherichia coli SSB protein has a similar but somewhat less robust capacity to displace a DNA strand annealed adjacent to a single-strand extension. These activities are likely to be relevant to the action of bacterial SSB proteins in double-strand break repair, acting at the frayed ends created by ionizing radiation. -
Crystal structure of Deinococcus radiodurans single-stranded DNA-binding protein suggests a mechanism for coping with DNA damage
Proceedings of the National Academy of Sciences U.S.A. 2004. 101(23):8575-80. Bernstein DA, Eggington JM, Killoran MP, Misic AM, Cox MM, Keck JL.
Abstract
Single-stranded DNA (ssDNA)-binding (SSB) proteins are uniformly required to bind and protect single-stranded intermediates in DNA metabolic pathways. All bacterial and eukaryotic SSB proteins studied to date oligomerize to assemble four copies of a conserved domain, called an oligonucleotide/oligosaccharide-binding (OB) fold, that cooperate in nonspecific ssDNA binding. The vast majority of bacterial SSB family members function as homotetramers, with each monomer contributing a…Abstract
Single-stranded DNA (ssDNA)-binding (SSB) proteins are uniformly required to bind and protect single-stranded intermediates in DNA metabolic pathways. All bacterial and eukaryotic SSB proteins studied to date oligomerize to assemble four copies of a conserved domain, called an oligonucleotide/oligosaccharide-binding (OB) fold, that cooperate in nonspecific ssDNA binding. The vast majority of bacterial SSB family members function as homotetramers, with each monomer contributing a single OB fold. However, SSB proteins from the Deinococcus-Thermus genera are exceptions to this rule, because they contain two OB folds per monomer. To investigate the structural consequences of this unusual arrangement, we have determined a 1.8-A-resolution x-ray structure of Deinococcus radiodurans SSB. The structure shows that D. radiodurans SSB comprises two OB domains linked by a beta-hairpin motif. The protein assembles a four-OB-fold arrangement by means of symmetric dimerization. In contrast to homotetrameric SSB proteins, asymmetry exists between the two OB folds of D. radiodurans SSB because of sequence differences between the domains. These differences appear to reflect specialized roles that have evolved for each domain. Extensive crystallographic contacts link D. radiodurans SSB dimers in an arrangement that has important implications for higher-order structures of the protein bound to ssDNA. This assembly utilizes the N-terminal OB domain and the beta-hairpin structure that is unique to Deinococcus and Thermus species SSB proteins. We hypothesize that differences between D. radiodurans SSB and homotetrameric bacterial SSB proteins may confer a selective advantage to D. radiodurans cells that aids viability in environments that challenge genomic stability. -
The single-stranded DNA-binding protein of Deinococcus radiodurans
BMC Microbiology. 2004. 12;4:2. Eggington JM, Haruta N, Wood EA, Cox MM.
Abstract
BACKGROUND: Deinococcus radiodurans R1 is one of the most radiation-resistant organisms known and is able to repair an unusually large amount of DNA damage without induced mutation. Single-stranded DNA-binding (SSB) protein is an essential protein in all organisms and is involved in DNA replication, recombination and repair. The published genomic sequence from Deinococcus radiodurans includes a putative single-stranded DNA-binding protein gene (ssb; DR0100) requiring a translational…Abstract
BACKGROUND: Deinococcus radiodurans R1 is one of the most radiation-resistant organisms known and is able to repair an unusually large amount of DNA damage without induced mutation. Single-stranded DNA-binding (SSB) protein is an essential protein in all organisms and is involved in DNA replication, recombination and repair. The published genomic sequence from Deinococcus radiodurans includes a putative single-stranded DNA-binding protein gene (ssb; DR0100) requiring a translational frameshift for synthesis of a complete SSB protein. The apparently tripartite gene has inspired considerable speculation in the literature about potentially novel frameshifting or RNA editing mechanisms. Immediately upstream of the ssb gene is another gene (DR0099) given an ssb-like annotation, but left unexplored.
RESULTS: ...
CONCLUSIONS: The correct Deinococcus radiodurans ssb gene is a contiguous open reading frame that codes for the largest bacterial SSB monomer identified to date. The Deinococcus radiodurans SSB protein includes two OB folds per monomer and functions as a homodimer. The Deinococcus radiodurans SSB protein efficiently stimulates Deinococcus radiodurans RecA and also Escherichia coli RecA protein-promoted DNA strand exchange reactions. The identification and purification of Deinococcus radiodurans SSB protein not only allows for greater understanding of the SSB protein family but provides an essential yet previously missing player in the current efforts to understand the extraordinary DNA repair capacity of Deinococcus radiodurans. -
A Convenient One-Step Gingerol Synthesis
Synthetic Communications. 1999. 29(11):1933-39. Fleming SA, Dyer CW, Eggington J.
Abstract
Racemic 6-gingerol can be obtained in a one-pot reaction by hexanal addition to the dianion of zingerone at low temperature. Similarly, addition of octanal or decanal to the dianion provides 8-gingerol or 10-gingerol, respectively. Acid treatment of the gingerols allows for formation of the corresponding shogaols.
Patents
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Method to Protect DNA Ends
Issued US 7,211,393
A method of protecting the 3' end of a DNA molecule from nuclease damage is disclosed. In one embodiment, the method comprises the step of exposing the DNA molecule to a preparation of DdrA protein.
Authors - Cox MM, Harris DR, Saveliev SV, Battista JR, Jolivet E, Tanaka M, Eggington JM
Courses
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How to Supervise & Produce Real Results
Management Training
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University of Utah Masters Level Statistics for Biomedical Informatics
BMI 6105
Projects
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Selected Public Speaking Engagements
- Present
TOO MANY TO CONTINUE LISTING
UPENN Cancer Risk Evaluation Program Case Conference - Invited Speaker.
Variant Classification.
Eggington J. Remote webcast to University of Pennsylvania, PA. 1/27/ 2017.
The US Oncology Network, Genetics Risk and Evaluation Testing Conference.
Technical Evaluation of Genetic Testing: Quality Measures and Variant Classification.
Eggington J. New Orleans, LA. 9/13/ 2014.
National Society of Genetic Counselors (NSGC) – 32nd Annual…TOO MANY TO CONTINUE LISTING
UPENN Cancer Risk Evaluation Program Case Conference - Invited Speaker.
Variant Classification.
Eggington J. Remote webcast to University of Pennsylvania, PA. 1/27/ 2017.
The US Oncology Network, Genetics Risk and Evaluation Testing Conference.
Technical Evaluation of Genetic Testing: Quality Measures and Variant Classification.
Eggington J. New Orleans, LA. 9/13/ 2014.
National Society of Genetic Counselors (NSGC) – 32nd Annual Education Conference (AEC). Hereditary Cancer Testing Update - Myriad Genetic Laboratories Variant Classification. Eggington J, Mancini-Dinardo D, Kerr I. Anaheim, CA. 10/11/2013.
NSGC – 32nd AEC. Variant of Uncertain Significance Classification in Highly-Penetrant Cancer Susceptibility Genes: Tools for Interpretation, Psychosocial issues and Decision Making Implications. Culver J, Richards S, Tavtigian S, Eggington J, Lindor N. Anaheim, CA. 10/10/2013.
European Society of Human Genetics Conference, Myriad Corporate Satellite Meeting. Designing Variant Classification for a Hereditary Cancer Panel. Paris, France. 6/9/2013.
NSGC – 30th AEC. Classification of Variants in Cancer Predisposition Genes. Eggington J, Bowling L. San Diego, CA. 10/30/2011.
NSGC – 30th AEC. Perplexing Variants of Uncertain Significance. Bennett RL, Eggington J, Ward PA, Wiyrick SJ. San Diego, CA. 10/28/2011.
Texas Society of Genetic Counselors – 4th AEC. How Laboratories Classify Genetic Changes, Implications for Clinical Practice. Houston, TX. 4/9/2011.
Challenges in Clinical Cancer Genetics Conference. Perplexing Variants of Uncertain Significance: Laboratory Perspective. U of Washington. Seattle, WA. 9/27/2010.
Cancer Genetics Education Program at City of Hope – Invited Speaker. Variant Classification & Myriad’s New Variant Classification Program. City of Hope. Duarte, CA. 7/9/2010.
Honors & Awards
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Sego Award Winner
Sego Awards - Utah's first awards for Female Founders & CEOs
Awarded 2nd place for Utah Founder/CEO in the "Innovation in Technology" category.
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“Top 25” Myriad Employee Award
Myriad Genetic Laboratories, Inc.
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Ruth L. Kirschstein National Research Service Award
National Institutes of Health
Stipend and institutional costs for 2 years of Postdoctoral Fellowship. (Awarded but declined due to prior competing fellowship acceptance).
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AFSP Postdoctoral Fellowship Award
American Foundation for Suicide Prevention
Postdoctoral stipend and institutional costs for two years.
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Steenbock Predoctoral Fellowship Award
University of Wisconsin-Madison
For research accomplishments. Full support, tuition and fees.
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Multiple “Cause for Applause” Awards
Medical Services, Myriad Genetic Laboratories, Inc.
2010 - 2012
For outstanding scientific presentations to clients and the forging of collaborations.
Recommendations received
15 people have recommended Julie
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