Abstract

The protein synthesis factor EF-1Alpha of Mucor racemosus hyphae contains 8 or 9 methylated lysines per molecule, while the factor from assexual sporangiospores is non-methylated. During the course of spore germination the specific acitivity increases 6 to 9-fold. This increase in specific activity is accompanied by a constant level of mRNA and a level of EF-1Alpha protein that changes little during the germination period. By isolating and sequencing the EF-1Alpha gene, we found tht it is present in 3 copies in the Mucor genome, each in a different genomic context. This proposal is concerned with two aspects of EF-1Alpha: the role of methylation in """"""""activating"""""""" the factor, and the genetic organization, including the control of expression of each gene. We will study the effect of methylation on various EF-1Alpha activities in vitro, locate each methylated site within the protein and purify and study the EF-1Alpha methylases. We will further prepare gene-specific hybridization probes to assess the expression of each gene during several morphogenetic changes (e.g., spore germination, yeast-to-hypha conversions, and sporulation). We also propose to study the 5'flanking regions of each gene to assess the mechanisms controlling transcription.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM023999-09
Application #
3271989
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1977-04-01
Project End
1991-11-30
Budget Start
1986-12-01
Budget End
1987-11-30
Support Year
9
Fiscal Year
1987
Total Cost
Indirect Cost

  Institution

Name
University of California Irvine
Department
Type
Schools of Medicine
DUNS #
161202122
City
Irvine
State
CA
Country
United States
Zip Code
92697

  Publications

Myers, K K; Fonzi, W A; Sypherd, P S (1992) Isolation and sequence analysis of the gene for translation elongation factor 3 from Candida albicans. Nucleic Acids Res 20:1705-10
Saporito, S M; Sypherd, P S (1991) The isolation and characterization of a calmodulin-encoding gene (CMD1) from the dimorphic fungus Candida albicans. Gene 106:43-9
Sundstrom, P; Irwin, M; Smith, D et al. (1991) Both genes for EF-1 alpha in Candida albicans are translated. Mol Microbiol 5:1703-6
Sundstrom, P; Smith, D; Sypherd, P S (1990) Sequence analysis and expression of the two genes for elongation factor 1 alpha from the dimorphic yeast Candida albicans. J Bacteriol 172:2036-45
Sherman, M; Sypherd, P S (1989) Role of lysine methylation in the activities of elongation factor 1 alpha. Arch Biochem Biophys 275:371-8
Sosa, L; Fonzi, W A; Sypherd, P S (1989) Structure of a ribosomal protein gene in Mucor racemosus. Nucleic Acids Res 17:9319-31
Fonzi, W A; Sypherd, P S (1987) The gene and the primary structure of ornithine decarboxylase from Saccharomyces cerevisiae. J Biol Chem 262:10127-33
Sundstrom, P; Lira, L M; Choi, D et al. (1987) Sequence analysis of the EF-1 alpha gene family of Mucor racemosus. Nucleic Acids Res 15:9997-10006
Linz, J E; Sypherd, P S (1987) Expression of three genes for elongation factor 1 alpha during morphogenesis of Mucor racemosus. Mol Cell Biol 7:1925-32
Linz, J E; Lira, L M; Sypherd, P S (1986) The primary structure and the functional domains of an elongation factor-1 alpha from Mucor racemosus. J Biol Chem 261:15022-9

Showing the most recent 10 out of 16 publications