Werner's syndrome (WS) is a homozygous recessive disease characterized by early onset of many characteristics of normal aging, such as wrinkling of the skin, graying of the hair, cataracts, diabetes, and osteoporosis. Because of the acceleration of aging in WS, the study of this disease will hopefully shed light on the degenerative processes that occur in normal aging. Cells from WS patients grow more slowly and senescence at an earlier population doubling than age-matched normal cells, possibly because these cells appear to lose the telomeric ends of their chromosomes at an accelerated rate. In general, WS cells have a high level of genomic instability, with increased amounts of DNA deletions, insertions, and rearrangements. These effects could potentially be the result of defects in DNA repair, replication, and/or recombination, although the actual biochemical defect remains unknown. The gene that is defective in WS, the WRN gene, has been identified and characterized. We have made purified WRN protein for use in a number of basic and complex biochemical assays. We are pursuing several avenues to identify and characterize the biochemical defect in WS cells. WRN protein has helicase activity and will unwind small and large DNA duplex constructs. Additionally, WRN has a 3-5'exonuclease function. Recently, we showed that the RQC domain of WRN is critical for its DNA binding and catalytic activities. This work also revealed that mutations in the RQC could impact the exonuclease domain of WRN, which was previously thought to be an autonomous domain. We are comparing WRN to that of the other RecQ helicases that are all involved in the maintenance of genome stability. We continue to use confocal microscopy as a means to investigate the dynamic behavior of WRN and the other RecQ helicases. Specifically, WRN is recruited to DNA damage either by protein-protein interactions or due to protein post-translational modifications (PMTs). We are exploring the recruitment and retention dynamics of WRN proteins carrying point mutations or with alterations within potential PMT sites to determine if these elements impact WRNs DNA damage recruitment. Future studies aim to characterize more fully the protein complexes that WRN participates in both before and after DNA damage.

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Investigator-Initiated Intramural Research Projects (ZIA)
Project #
1ZIAAG000721-05
Application #
8736596
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
5
Fiscal Year
2013
Total Cost
$591,717
Indirect Cost
Name
National Institute on Aging
Department
Type
DUNS #
City
State
Country
Zip Code
Shamanna, Raghavendra A; Lu, Huiming; Croteau, Deborah L et al. (2016) Camptothecin targets WRN protein: mechanism and relevance in clinical breast cancer. Oncotarget 7:13269-84
Khadka, Prabhat; Croteau, Deborah L; Bohr, Vilhelm A (2016) RECQL5 has unique strand annealing properties relative to the other human RecQ helicase proteins. DNA Repair (Amst) 37:53-66
Tippana, Ramreddy; Hwang, Helen; Opresko, Patricia L et al. (2016) Single-molecule imaging reveals a common mechanism shared by G-quadruplex-resolving helicases. Proc Natl Acad Sci U S A 113:8448-53
Shin, Soochul; Lee, Jinwoo; Yoo, Sangwoon et al. (2016) Active Control of Repetitive Structural Transitions between Replication Forks and Holliday Junctions by Werner Syndrome Helicase. Structure 24:1292-300
Khadka, Prabhat; Hsu, Joseph K; Veith, Sebastian et al. (2015) Differential and Concordant Roles for Poly(ADP-Ribose) Polymerase 1 and Poly(ADP-Ribose) in Regulating WRN and RECQL5 Activities. Mol Cell Biol 35:3974-89
Edwards, Deanna N; Machwe, Amrita; Chen, Li et al. (2015) The DNA structure and sequence preferences of WRN underlie its function in telomeric recombination events. Nat Commun 6:8331
Shamanna, Raghavendra A; Singh, Dharmendra Kumar; Lu, Huiming et al. (2014) RECQ helicase RECQL4 participates in non-homologous end joining and interacts with the Ku complex. Carcinogenesis 35:2415-24
Bendtsen, Kristian Moss; Jensen, Martin Borch; May, Alfred et al. (2014) Dynamics of the DNA repair proteins WRN and BLM in the nucleoplasm and nucleoli. Eur Biophys J :
Debrabant, Birgit; Soerensen, Mette; Flachsbart, Friederike et al. (2014) Human longevity and variation in DNA damage response and repair: study of the contribution of sub-processes using competitive gene-set analysis. Eur J Hum Genet 22:1131-6
Lu, H; Fang, E F; Sykora, P et al. (2014) Senescence induced by RECQL4 dysfunction contributes to Rothmund-Thomson syndrome features in mice. Cell Death Dis 5:e1226

Showing the most recent 10 out of 50 publications