Abstract

We are studying how newly-made non-cytoplasmic proteins are sorted to their many distinct locations within cells. This involves protein assembly into membranes and selective protein transfer across membranes (secretion). Such non-cytoplasmic proteins are often made with amino-terminal leader sequences; we have purified the first membrane protein precursor with an intact leader sequence (M13 procoat) and the first """"""""leader peptidase"""""""" which removes the leader. Our reconstitution of assembly with these pure components will allow us to study at a chemical level how polar regions of polypeptide cross the bilayer. The structure, biogenesis, and specificity of the leader peptidase, as well as the fate of the leader peptide, will be examined. Assays for new elements which may function in assembly will be developed. Mutants in leader peptidase which are being isolated in our lab will allow a determination of the spatial distribution of assembly intermediates.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM023377-10
Application #
3271612
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1976-05-01
Project End
1988-06-30
Budget Start
1985-07-01
Budget End
1986-06-30
Support Year
10
Fiscal Year
1985
Total Cost
Indirect Cost

  Institution

Name
University of California Los Angeles
Department
Type
Schools of Medicine
DUNS #
119132785
City
Los Angeles
State
CA
Country
United States
Zip Code
90095

  Publications

Song, Hongki; Wickner, William (2017) A short region upstream of the yeast vacuolar Qa-SNARE heptad-repeats promotes membrane fusion through enhanced SNARE complex assembly. Mol Biol Cell 28:2282-2289
Song, Hongki; Orr, Amy; Duan, Mengtong et al. (2017) Sec17/Sec18 act twice, enhancing membrane fusion and then disassembling cis-SNARE complexes. Elife 6:
Schwartz, Matthew L; Nickerson, Daniel P; Lobingier, Braden T et al. (2017) Sec17 (?-SNAP) and an SM-tethering complex regulate the outcome of SNARE zippering in vitro and in vivo. Elife 6:
Orr, Amy; Song, Hongki; Rusin, Scott F et al. (2017) HOPS catalyzes the interdependent assembly of each vacuolar SNARE into a SNARE complex. Mol Biol Cell 28:975-983
Zick, Michael; Wickner, William (2016) Improved reconstitution of yeast vacuole fusion with physiological SNARE concentrations reveals an asymmetric Rab(GTP) requirement. Mol Biol Cell 27:2590-7
Zick, Michael; Orr, Amy; Schwartz, Matthew L et al. (2015) Sec17 can trigger fusion of trans-SNARE paired membranes without Sec18. Proc Natl Acad Sci U S A 112:E2290-7
Orr, Amy; Wickner, William; Rusin, Scott F et al. (2015) Yeast vacuolar HOPS, regulated by its kinase, exploits affinities for acidic lipids and Rab:GTP for membrane binding and to catalyze tethering and fusion. Mol Biol Cell 26:305-15
Baker, Richard W; Jeffrey, Philip D; Zick, Michael et al. (2015) A direct role for the Sec1/Munc18-family protein Vps33 as a template for SNARE assembly. Science 349:1111-4
Zick, Michael; Wickner, William T (2014) A distinct tethering step is vital for vacuole membrane fusion. Elife 3:e03251
Zick, Michael; Stroupe, Christopher; Orr, Amy et al. (2014) Membranes linked by trans-SNARE complexes require lipids prone to non-bilayer structure for progression to fusion. Elife 3:e01879

Showing the most recent 10 out of 39 publications