Abstract

Transposable elements play a major role in mediating DNA rearrangements within and between prokaryotic organisms. We propose to investigate the mechanism and regulation of transposition by prokaryotic insertion sequences. Our analysis will particularly emphasize transposon Tn10 and its component IS10 sequences, but will also analyze some aspects of IS26 and IS5. Seven specific projects are proposed below: (1) Biochemical analysis of IS10 transposition in vitro. (2) Genetic and physical analysis of IS10 transposase protein: isolation and characterization of general and specific classes of transposase mutants. (3) Genetic and biochemical comparison of transposition by IS26 and IS5 to that of IS10. (4) Genetic analysis of the fate of the transposon donor molecule during transposition. (5) Dissection of IS10 insertion specificity and investigation of the role of DNA melting in regional specificity of IS10 and IS26. (6) Mechanism of IS10-mediated translational control by anti-sense RNA: in vitro analysis of pairing between RNA-OUT and RNA-IN and binding of ribosomes to paired and unpaired RNAs. (7) Regulation of IS10 transposition by dam methylation: kinetics of methylation on non-replicating and replicating DNA in vivo and in vitro.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM025326-10
Application #
3272929
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1978-07-01
Project End
1991-06-30
Budget Start
1987-07-01
Budget End
1988-06-30
Support Year
10
Fiscal Year
1987
Total Cost
Indirect Cost

  Institution

Name
Harvard University
Department
Type
Schools of Arts and Sciences
DUNS #
071723621
City
Cambridge
State
MA
Country
United States
Zip Code
02138

  Publications

Kleckner, Nancy E; Chatzi, Katerina; White, Martin A et al. (2018) Coordination of Growth, Chromosome Replication/Segregation, and Cell Division in E. coli. Front Microbiol 9:1469
Gutu, Andrian; Chang, Frederick; O'Shea, Erin K (2018) Dynamical localization of a thylakoid membrane binding protein is required for acquisition of photosynthetic competency. Mol Microbiol 108:16-31
Liu, Chenli; Danilowicz, Claudia; Kleckner, Nancy et al. (2017) Single molecule identification of homology-dependent interactions between long ssRNA and dsDNA. Nucleic Acids Res 45:894-901
Gladyshev, Eugene; Kleckner, Nancy (2017) Recombination-independent recognition of DNA homology for repeat-induced point mutation. Curr Genet 63:389-400
Gladyshev, Eugene; Kleckner, Nancy (2016) Recombination-Independent Recognition of DNA Homology for Repeat-Induced Point Mutation (RIP) Is Modulated by the Underlying Nucleotide Sequence. PLoS Genet 12:e1006015
Yoon, Sang-Wook; Lee, Min-Su; Xaver, Martin et al. (2016) Meiotic prophase roles of Rec8 in crossover recombination and chromosome structure. Nucleic Acids Res 44:9296-9314
Zheng, Hai; Ho, Po-Yi; Jiang, Meiling et al. (2016) Interrogating the Escherichia coli cell cycle by cell dimension perturbations. Proc Natl Acad Sci U S A 113:15000-15005
Kleckner, Nancy (2016) Questions and Assays. Genetics 204:1343-1349
Yang, Darren; Boyer, Benjamin; Prévost, Chantal et al. (2015) Integrating multi-scale data on homologous recombination into a new recognition mechanism based on simulations of the RecA-ssDNA/dsDNA structure. Nucleic Acids Res 43:10251-63
Peacock-Villada, Alexandra; Coljee, Vincent; Danilowicz, Claudia et al. (2015) ssDNA Pairing Accuracy Increases When Abasic Sites Divide Nucleotides into Small Groups. PLoS One 10:e0130875

Showing the most recent 10 out of 83 publications