Abstract

The goal of the project is to understand the intramolecular interactions involved in autocatalytic RNA splicing, and the role of splicing in the regulatory biology of a simple organism, bacteriophage T4. Two new introns have been discovered in T4, and have localized near genes nrdB and nrdC. These introns will be characterized with respect to splicing intermediates, cofactors, and nucleotide sequence. Comparisons will be made with the characterized td intron of T4 and with other Class I introns of eucaryotes. Insertional mutagenesis of the introns and their flanking sequences, with a Beta-galactosidase fusion gene, will allow determination of the genes which contain the introns. Selection schemes have been devised to find mutations that block splicing, as well as mutations at a second site which reactivate it. Insertion of introns into the rIIB gene will provide a sensitive biological assay for splicing. The distribution of mutable sites in the introns will be analyzed to determine the interactions that contribute to the autocatalytic splicing mechanism. Also, a novel hypothesis, which suggests that the splicing reaction is coupled to a metabolic regulatory circuit, will be tested.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
1R01GM037746-01
Application #
3293392
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1987-03-01
Project End
1990-02-28
Budget Start
1987-03-01
Budget End
1988-02-29
Support Year
1
Fiscal Year
1987
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
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
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
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