Katherine L. (Kathy) Friedman, Ph.D.
Associate Professor of Biological Sciences
Vanderbilt Biological Sciences Department
2417A Stevenson Ctr, 1161 21st Av S
Nashville, TN 37235
Our laboratory is interested in mechanisms that maintain genome stability, with research endeavors that lie at the intersection of DNA replication and DNA repair.
The Friedman lab is actively recruiting rotation students! Please contact Dr. Friedman for more information.
Telomeres: The ends of chromosomes (telomeres) constitute a tiny fraction of DNA in a cell, but play a central role in genome stability and cellular lifespan. Processing of the leading strand results in chromosome shortening following conventional replication. A ribonucleoprotein enzyme, telomerase, replenishes these terminal sequences. Since the catalytic subunit of telomerase is not expressed in most human somatic cells, human chromosomes lose telomere sequence upon successive cell divisions, a process that limits proliferative capacity. DNA loss is reversed in the vast majority of cancers by reactivation of telomerase, allowing cells to far exceed their normal life expectancy. Since inhibition of telomerase triggers apoptosis or senescence of some tumor cell lines, telomerase is a promising target of anti-cancer therapy. Genetic experiments initiated two decades ago established the yeast Saccharomyces cerevisiae as a key model system for studies of telomerase. Furthermore, conservation of several telomerase components between yeast and human suggests that many aspects of yeast telomere biology have correlates in human cells. The ease with which the cell cycle can be manipulated and the vast array of genetic approaches make yeast an ideal system in which to study mechanisms and consequences of telomerase regulation at the cellular level.
Telomerase and double-strand breaks: Telomerase has the ability to create a new, functional telomere at an internal site following a DSB. Such de novo telomere addition can interfere with normal DNA repair through the creation of terminal deletions that may or may not be compatible with cell viability. Chromosome truncations, some of which arise through de novo telomere addition, are a common source of human genetic disease. Given these consequences, it is not surprising that cells have evolved mechanisms to inhibit the action of telomerase at DSBs. However, work recently published from our laboratory characterized two sequences in the yeast genome that support much higher frequencies of de novo telomere addition than neighboring regions [termed SIRTAs (Sites of Internal Repair-associated Telomere Addition)]. Such sites have the potential to generate genomic instability through the generation of terminal deletions, but may provide a novel mechanism of DNA damage tolerance by preventing chromosome loss in the face of an otherwise unrepaired break.
In current work, we are testing the hypothesis that specific cis- and trans-acting factors render SiRTAs refractory to mechanisms that normally inhibit telomerase action at a DSB and examining the possibility that telomerase action is differentially regulated when a break has persisted without repair. We are identifying additional SiRTAs to facilitate comparative analyses and to determine whether SiRTAs are non-randomly distributed in the genome as would be predicted if such sites impact genome stability. Genetic screening is ongoing to identify factors that contribute to telomere addition at SiRTAs. The high frequency of telomere addition at a single site make SiRTAs excellent models to elucidate the mechanisms through which crucial decisions regarding DSB repair pathway choice and efficiency are achieved.
Mechanisms of DNA double-strand break repair: Generation of a single double-strand break (DSB) through expression of a site-specific nuclease in yeast has been instrumental in our understanding of DNA damage repair mechanisms. However, these systems are limited by the requirement for a specific recognition site (precluding experiments to examine sequence context immediately surrounding the break) and by the generation of non-natural staggered breaks in DNA. The CRISPR/Cas9 system generates a blunt (or nearly blunt) DSB that can be targeted to a nearly limitless number of endogenous sequences in the genome. This system reveals functions during the initial stages of DNA end processing and repair that are different from those observed using previous experimental paradigms. We aim to characterize the kinetics of DNA repair that occur following CRISPR/Cas9 cleavage. Specifically, we are pursuing the contribution of the Rif1 protein and the Pol4 DNA polymerase to the early steps of DNA DSB repair. In addition to providing insight into basic mechanisms of DNA repair, these experiments will generate significant information about how CRISPR/Cas9 interacts with its chromosomal target site and with the repair machinery, knowledge that will inform the use of this system for genomic engineering in mammalian cells.
- Schaller MD, McDowell G, Porter A, Shippen D, Friedman KL, Gentry MS, Serio TR, Sundquist WI. What's in a name?. Elife. 2017 Oct 10/24/2017; 6: PMID: 29063834, PMCID: PMC5655148, DOI: 10.7554/eLife.32437, ISSN: 2050-084X.
Available from: http://www.ncbi.nlm.nih.gov/pubmed/29063834.
- Obodo UC, Epum EA, Platts MH, Seloff J, Dahlson NA, Velkovsky SM, Paul SR, Friedman KL. Endogenous Hot Spots of De Novo Telomere Addition in the Yeast Genome Contain Proximal Enhancers That Bind Cdc13. Mol. Cell. Biol [electronic-print]. 2016 Jun 6/15/2016; 36(12): 1750-63. PMID: 27044869, PMCID: PMC4907100, PII: MCB.00095-16, DOI: 10.1128/MCB.00095-16, ISSN: 1098-5549.
Available from: http://www.ncbi.nlm.nih.gov/pubmed/27044869.
- Ning B, Feldkamp MD, Cortez D, Chazin WJ, Friedman KL, Fanning E. Simian virus Large T antigen interacts with the N-terminal domain of the 70 kD subunit of Replication Protein A in the same mode as multiple DNA damage response factors. PLoS ONE. 2015; 10(2): e0116093. PMID: 25706313, PMCID: PMC4337903, PII: PONE-D-14-45260, DOI: 10.1371/journal.pone.0116093, ISSN: 1932-6203.
Available from: http://www.ncbi.nlm.nih.gov/pubmed/25706313.
- Sowd GA, Mody D, Eggold J, Cortez D, Friedman KL, Fanning E. SV40 utilizes ATM kinase activity to prevent non-homologous end joining of broken viral DNA replication products. PLoS Pathog. 2014 Dec; 10(12): e1004536. PMID: 25474690, PMCID: PMC4256475, PII: PPATHOGENS-D-14-01844, DOI: 10.1371/journal.ppat.1004536, ISSN: 1553-7374.
Available from: http://www.ncbi.nlm.nih.gov/pubmed/25474690.
- Hawkins C, Friedman KL. Normal telomere length maintenance in Saccharomyces cerevisiae requires nuclear import of the ever shorter telomeres 1 (Est1) protein via the importin alpha pathway. Eukaryotic Cell [print-electronic]. 2014 Aug; 13(8): 1036-50. PMID: 24906415, PMCID: PMC4135794, PII: EC.00115-14, DOI: 10.1128/EC.00115-14, ISSN: 1535-9786.
Available from: http://www.ncbi.nlm.nih.gov/pubmed/24906415.
- Ferguson JL, Chao WC, Lee E, Friedman KL. The anaphase promoting complex contributes to the degradation of the S. cerevisiae telomerase recruitment subunit Est1p. PLoS ONE [print-electronic]. 2013; 8(1): e55055. PMID: 23372810, PMCID: PMC3555863, PII: PONE-D-12-35468, DOI: 10.1371/journal.pone.0055055, ISSN: 1932-6203.
Available from: http://www.ncbi.nlm.nih.gov/pubmed/23372810.
- Bairley RC, Guillaume G, Vega LR, Friedman KL. A mutation in the catalytic subunit of yeast telomerase alters primer-template alignment while promoting processivity and protein-DNA binding. J. Cell. Sci [print-electronic]. 2011 Dec 12/15/2011; 124(Pt 24): 4241-52. PMID: 22193961, PMCID: PMC4074303, PII: jcs.090761, DOI: 10.1242/jcs.090761, ISSN: 1477-9137.
Available from: http://www.ncbi.nlm.nih.gov/pubmed/22193961.
- Talley JM, DeZwaan DC, Maness LD, Freeman BC, Friedman KL. Stimulation of yeast telomerase activity by the ever shorter telomere 3 (Est3) subunit is dependent on direct interaction with the catalytic protein Est2. J. Biol. Chem [print-electronic]. 2011 Jul 7/29/2011; 286(30): 26431-9. PMID: 21659533, PMCID: PMC3143607, PII: M111.228635, DOI: 10.1074/jbc.M111.228635, ISSN: 1083-351X.
Available from: http://www.ncbi.nlm.nih.gov/pubmed/21659533.
- Friedman KL. Telomerase reverse transcriptase and Wnt signaling. Mol. Cell. Biol [print-electronic]. 2011 Jun; 31(12): 2366-8. PMID: 21536649, PMCID: PMC3133428, PII: MCB.05462-11, DOI: 10.1128/MCB.05462-11, ISSN: 1098-5549.
Available from: http://www.ncbi.nlm.nih.gov/pubmed/21536649.
- Osterhage JL, Friedman KL. Chromosome end maintenance by telomerase. J. Biol. Chem [print-electronic]. 2009 Jun 6/12/2009; 284(24): 16061-5. PMID: 19286666, PMCID: PMC2713563, PII: R900011200, DOI: 10.1074/jbc.R900011200, ISSN: 0021-9258.
Available from: http://www.ncbi.nlm.nih.gov/pubmed/19286666.
- Ji H, Adkins CJ, Cartwright BR, Friedman KL. Yeast Est2p affects telomere length by influencing association of Rap1p with telomeric chromatin. Mol. Cell. Biol [print-electronic]. 2008 Apr; 28(7): 2380-90. PMID: 18212041, PMCID: PMC2268414, PII: MCB.01648-07, DOI: 10.1128/MCB.01648-07, ISSN: 1098-5549.
Available from: http://www.ncbi.nlm.nih.gov/pubmed/18212041.
- Osterhage JL, Talley JM, Friedman KL. Proteasome-dependent degradation of Est1p regulates the cell cycle-restricted assembly of telomerase in Saccharomyces cerevisiae. Nat. Struct. Mol. Biol [print-electronic]. 2006 Aug; 13(8): 720-8. PMID: 16862158, PII: nsmb1125, DOI: 10.1038/nsmb1125, ISSN: 1545-9993.
Available from: http://www.ncbi.nlm.nih.gov/pubmed/16862158.
- Ji H, Platts MH, Dharamsi LM, Friedman KL. Regulation of telomere length by an N-terminal region of the yeast telomerase reverse transcriptase. Mol. Cell. Biol. 2005 Oct; 25(20): 9103-14. PMID: 16199886, PMCID: PMC1265764, PII: 25/20/9103, DOI: 10.1128/MCB.25.20.9103-9114.2005, ISSN: 0270-7306.
Available from: http://www.ncbi.nlm.nih.gov/pubmed/16199886.
- Friedman KL, Heit JJ, Long DM, Cech TR. N-terminal domain of yeast telomerase reverse transcriptase: recruitment of Est3p to the telomerase complex. Mol. Biol. Cell. 2003 Jan; 14(1): 1-13. PMID: 12529422, PMCID: PMC140223, DOI: 10.1091/mbc.E02-06-0327, ISSN: 1059-1524.
Available from: http://www.ncbi.nlm.nih.gov/pubmed/12529422.
- Friedman KL, Cech TR. Essential functions of amino-terminal domains in the yeast telomerase catalytic subunit revealed by selection for viable mutants. Genes Dev. 1999 Nov 11/1/1999; 13(21): 2863-74. PMID: 10557213, PMCID: PMC317136, ISSN: 0890-9369.
Available from: http://www.ncbi.nlm.nih.gov/pubmed/10557213.