Abstract

The overall goal of the research is to identify and characterize mutants and genes affecting structure and function in the nucleus, using S. cerevisiae as a model organism. There are three specific aims. The first is to use the two hybrid system to identify proteins that interact with DNA topoisomerases I and II and to characterize the in vivo roles of these proteins through standard genetic methods. The second is to continue studies on transcriptional silencing in yeast. SIR1, SIR3, and SIR4 homologs will be isolated from related yeasts, and, if possible, from S. pombe and metazoans to determine whether silencing by Sir proteins is evolutionarily conserved. The domains required for Sir1 functions in silencing will be identified. The role of S phase in the establishment of silencing by Sir1 will be determined, using as an assay Sir1 tethered to a silencer from which the ARS element has been deleted. Two-hybrid screens will be used with SIR1 and the N-terminus of SIR3 to identify interacting proteins. The third specific aim is to find mutants defective in the maintenance of nuclear integrity using a the fluorescent dye DiOC6 and to isolate the corresponding genes. Nuclear lamin genes in budding and fission yeast will be identified by screening for mutants that depend on the expression of the chicken lamin B gene for viability. A similar genetic approach will be applied to SIR3 and SIR4 to identify functionally equivalent genes that may participate in the attachment of telomeres to the nuclear periphery.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM028220-19
Application #
2734424
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1980-07-01
Project End
1999-06-30
Budget Start
1998-07-01
Budget End
1999-06-30
Support Year
19
Fiscal Year
1998
Total Cost
Indirect Cost

  Institution

Name
State University New York Stony Brook
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
804878247
City
Stony Brook
State
NY
Country
United States
Zip Code
11794

  Publications

Hsu, Hao-Chi; Wang, Chia-Lin; Wang, Mingzhu et al. (2013) Structural basis for allosteric stimulation of Sir2 activity by Sir4 binding. Genes Dev 27:64-73
Ren, Jie; Wang, Chia-Lin; Sternglanz, Rolf (2010) Promoter strength influences the S phase requirement for establishment of silencing at the Saccharomyces cerevisiae silent mating type Loci. Genetics 186:551-60
Sampath, Vinaya; Yuan, Peihua; Wang, Isabel X et al. (2009) Mutational analysis of the Sir3 BAH domain reveals multiple points of interaction with nucleosomes. Mol Cell Biol 29:2532-45
Krichevsky, Alexander; Gutgarts, Helen; Kozlovsky, Stanislav V et al. (2007) C2H2 zinc finger-SET histone methyltransferase is a plant-specific chromatin modifier. Dev Biol 303:259-69
Vaquero, Alejandro; Scher, Michael B; Lee, Dong Hoon et al. (2006) SirT2 is a histone deacetylase with preference for histone H4 Lys 16 during mitosis. Genes Dev 20:1256-61
Connelly, Jessica J; Yuan, Peihua; Hsu, Hao-Chi et al. (2006) Structure and function of the Saccharomyces cerevisiae Sir3 BAH domain. Mol Cell Biol 26:3256-65
Zappulla, David C; Maharaj, Arindel S R; Connelly, Jessica J et al. (2006) Rtt107/Esc4 binds silent chromatin and DNA repair proteins using different BRCT motifs. BMC Mol Biol 7:40
Xiang, Song; Kim, Eun Young; Connelly, Jessica J et al. (2006) The crystal structure of Cdc42 in complex with collybistin II, a gephyrin-interacting guanine nucleotide exchange factor. J Mol Biol 359:35-46
Liu, Bingsheng; Sutton, Ann; Sternglanz, Rolf (2005) A yeast polyamine acetyltransferase. J Biol Chem 280:16659-64
Wang, Xiaorong; Connelly, Jessica J; Wang, Chia-Lin et al. (2004) Importance of the Sir3 N terminus and its acetylation for yeast transcriptional silencing. Genetics 168:547-51

Showing the most recent 10 out of 35 publications