Abstract

This proposal will explore the possible involvement of RNA self-splicing in genetic regulatory networks of bacteriophage. 1. In vitro splicing assays will be carried out to determine if guanosine-like molecules, that are found in bacterial cells, can compete for the ribozyme active site and inhibit splicing. 2. Splicing assays will also be carried out in vivo, under a variety of conditions of cell growth, to determine if anaerobiosis, unbalanced growth, or physiological stress can result in modulation of the splicing reaction. 3. The organization of the phage introns suggests that there may be coupling between translation of the genes contained within the introns, and relative splicing efficiency. This model will be tested by in vitro mutagenesis of the introns (to produce various levels of translation), to test whether there is an inverse relationship between splicing and translation. 4. A combination of classical and molecular genetic methods will be applied to selection of intron mutations and second site revertants. This method will be used to determine the molecular interactions that determine splice site specificity.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM037746-06
Application #
3293397
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1987-03-01
Project End
1995-02-28
Budget Start
1992-03-01
Budget End
1993-02-28
Support Year
6
Fiscal Year
1992
Total Cost
Indirect Cost

  Institution

Name
State University of New York at Albany
Department
Type
Schools of Arts and Sciences
DUNS #
City
Albany
State
NY
Country
United States
Zip Code
12222

  Publications

Liu, Qingqing; Derbyshire, Victoria; Belfort, Marlene et al. (2006) Distance determination by GIY-YIG intron endonucleases: discrimination between repression and cleavage functions. Nucleic Acids Res 34:1755-64
Landthaler, Markus; Shen, Betty W; Stoddard, Barry L et al. (2006) I-BasI and I-HmuI: two phage intron-encoded endonucleases with homologous DNA recognition sequences but distinct DNA specificities. J Mol Biol 358:1137-51
Edgell, David R; Derbyshire, Victoria; Van Roey, Patrick et al. (2004) Intron-encoded homing endonuclease I-TevI also functions as a transcriptional autorepressor. Nat Struct Mol Biol 11:936-44
Shen, Betty W; Landthaler, Markus; Shub, David A et al. (2004) DNA binding and cleavage by the HNH homing endonuclease I-HmuI. J Mol Biol 342:43-56
Landthaler, Markus; Lau, Nelson C; Shub, David A (2004) Group I intron homing in Bacillus phages SPO1 and SP82: a gene conversion event initiated by a nicking homing endonuclease. J Bacteriol 186:4307-14
Landthaler, Markus; Shub, David A (2003) The nicking homing endonuclease I-BasI is encoded by a group I intron in the DNA polymerase gene of the Bacillus thuringiensis phage Bastille. Nucleic Acids Res 31:3071-7
Liu, Qingqing; Belle, Archana; Shub, David A et al. (2003) SegG endonuclease promotes marker exclusion and mediates co-conversion from a distant cleavage site. J Mol Biol 334:13-23
Belle, Archana; Landthaler, Markus; Shub, David A (2002) Intronless homing: site-specific endonuclease SegF of bacteriophage T4 mediates localized marker exclusion analogous to homing endonucleases of group I introns. Genes Dev 16:351-62
Landthaler, Markus; Begley, Ulrike; Lau, Nelson C et al. (2002) Two self-splicing group I introns in the ribonucleotide reductase large subunit gene of Staphylococcus aureus phage Twort. Nucleic Acids Res 30:1935-43
Bonocora, R P; Shub, D A (2001) A novel group I intron-encoded endonuclease specific for the anticodon region of tRNA(fMet) genes. Mol Microbiol 39:1299-306

Showing the most recent 10 out of 31 publications