Research in the Keck lab examines the structural mechanisms that drive DNA replication, replication restart, recombination, and repair reactions. Successful execution of these pathways is essential in all cells, and defects in the proteins that facilitate genome maintenance reactions lead to genome instability, cell death, and disease. Our studies combine structural approaches with biochemical and cell biological methods to answer fundamental structure-function questions in genome biology.
Single-stranded DNA-binding proteins. Bacterial single-stranded (ss) DNA-binding proteins (SSBs) play essential protective and organizational roles in genome biology. In their protective functions, SSBs bind and sequester ssDNA intermediates that are formed during genome maintenance reactions. As organizational centers, SSB/ssDNA complexes form dynamic protein-docking “hubs” at which over a dozen different DNA replication, recombination, and repair enzymes gain access to genomic substrates through direct interactions with SSB. This clustering of enzymes is thought to help integrate cellular genome maintenance reactions by facilitating the exchange of ssDNA substrates between DNA replication, recombination, and repair pathways. In all cases examined to date, the last ~6 residues of SSB’s flexible C-terminus (SSB-Ct) form its protein docking site. Eukaryotic SSBs also interact with a diverse array of genome maintenance proteins but, since they lack the SSB-Ct element found in bacterial SSBs, they do so through distinct mechanisms.
Honors & Awards
• Jane Coffin Childs Memorial Fund Postdoctoral Fellow, 1997-2000
• Shaw Scientist Award, 2003
• American Cancer Society Research Scholar, 2006
• WARF Romnes Faculty Fellowship, 2009
• School of Medicine and Public Health Dean’s Teaching Award, 2010
• UW Health Community Service Award 2013
• Liu K, Myers AR, Pisithkul T, Claas KR, Satyshur KA, Amador-Noguez D, Keck JL,
Wang JD. Molecular Mechanism and Evolution of Guanylate Kinase Regulation by
(p)ppGpp. Mol Cell. 2015 Feb 19;57(4):735-49. doi: 10.1016/j.molcel.2014.12.037.
Epub 2015 Feb 5. PubMed PMID: 25661490; PubMed Central PMCID: PMC4336630.
• Bhattacharyya B, Keck JL. Grip it and rip it: structural mechanisms of DNA
helicase substrate binding and unwinding. Protein Sci. 2014 Nov;23(11):1498-507.
doi: 10.1002/pro.2533. Epub 2014 Aug 22. PubMed PMID: 25131811; PubMed Central
• Walsh BW, Bolz SA, Wessel SR, Schroeder JW, Keck JL, Simmons LA. RecD2
helicase limits replication fork stress in Bacillus subtilis. J Bacteriol. 2014
Apr;196(7):1359-68. doi: 10.1128/JB.01475-13. Epub 2014 Jan 17. PubMed PMID:
24443534; PubMed Central PMCID: PMC3993351.
• Bhattacharyya B, George NP, Thurmes TM, Zhou R, Jani N, Wessel SR, Sandler SJ,
Ha T, Keck JL. Structural mechanisms of PriA-mediated DNA replication restart.
Proc Natl Acad Sci U S A. 2014 Jan 28;111(4):1373-8. doi:
10.1073/pnas.1318001111. Epub 2013 Dec 30. PubMed PMID: 24379377; PubMed Central
• Wessel SR, Marceau AH, Massoni SC, Zhou R, Ha T, Sandler SJ, Keck JL.
PriC-mediated DNA replication restart requires PriC complex formation with the
single-stranded DNA-binding protein. J Biol Chem. 2013 Jun 14;288(24):17569-78.
doi: 10.1074/jbc.M113.478156. Epub 2013 Apr 29. PubMed PMID: 23629733; PubMed
Central PMCID: PMC3682556.