Genetic Analysis of Protein Transport Into Mitochondria

Young, Elton T.

Abstract

The role of the signal sequence in targeting a protein to its intracellular location will be studied by genetic, biochemical, and recombinant DNA methods. By using the yeast Saccharomyces we can take a genetic approach to identifying both elements of structure of the leader itself, and the cellular components with which it interacts. Mutants will be isolated by virtue of being unable to correctly localize a protein synthesized in the cytoplasm and destined for the mitochondria, or by virtue of being inviable due to a defect in protein targeting. The structure of a synthetic mitochondrial targeting sequence will be determined by two- dimensional nuclear magnetic resonance spectroscopy. Mutant presequences, defective in import or cleavage will be studied to correlate alterations in structure with alterations in function. The synthetic presequence will also be used as a probe to try to identify proteins on the mitochondria that act as receptor for targeting proteins to that organelle.

Funding Agency

• Agency: National Institute of Health (NIH)
• Institute: National Institute of General Medical Sciences (NIGMS)
• Type: Research Project (R01)
• Project #: 5R01GM033779-08
• Application #: 3283778
• Study Section: Microbial Physiology and Genetics Subcommittee 2 (MBC)
• Project Start: 1984-07-01
• Project End: 1993-06-30
• Budget Start: 1991-07-01
• Budget End: 1992-06-30
• Support Year: 8
• Fiscal Year: 1991

Institution

• Name: University of Washington
• Type: Schools of Medicine
• DUNS #: 135646524
• City: Seattle
• State: WA
• Country: United States
• Zip Code: 98195

Publications

Cheng, C; Kacherovsky, N; Dombek, K M et al. (1994) Identification of potential target genes for Adr1p through characterization of essential nucleotides in UAS1. Mol Cell Biol 14:3842-52

Dombek, K M; Camier, S; Young, E T (1993) ADH2 expression is repressed by REG1 independently of mutations that alter the phosphorylation of the yeast transcription factor ADR1. Mol Cell Biol 13:4391-9

Mooney, D T; Pilgrim, D B; Young, E T (1990) Mutant alcohol dehydrogenase (ADH III) presequences that affect both in vitro mitochondrial import and in vitro processing by the matrix protease. Mol Cell Biol 10:2801-8

Allison, D S; Young, E T (1989) Mutations in the signal sequence of prepro-alpha-factor inhibit both translocation into the endoplasmic reticulum and processing by signal peptidase in yeast cells. Mol Cell Biol 9:4977-85

Allison, D S; Young, E T (1988) Single-amino-acid substitutions within the signal sequence of yeast prepro-alpha-factor affect membrane translocation. Mol Cell Biol 8:1915-22

Pilgrim, D; Young, E T (1987) Primary structure requirements for correct sorting of the yeast mitochondrial protein ADH III to the yeast mitochondrial matrix space. Mol Cell Biol 7:294-304