Abstract

The overall goal of this research is to develop a mechanistic understanding of the vacuolar biogenesis pathways in the yeast Saccharomyces cerevisiae. Genetic analysis has led to the identification of a very large number of components required along the vacuolar assembly pathway in yeast. A complex of proteins called GARP has been identified that functions in retrograde membrane transport to the late-Golgi compartment, and this complex will be investigated further both genetically and biochemically. GARP is one of the simplest and best characterized tethering complexes, and a complete molecular, biochemical and structural analysis of GARP should provide considerable insight into the role of tethering complexes in vesicle fusion. A genome-wide screen for new VPS genes has identified over 20 new genes/proteins involved in vacuolar biogenesis, and a number of these genes will be investigated further. Eight of these new Vps proteins are involved in transport steps between the Golgi and endosome, and will be investigated by genetic and biochemical approaches. Four of these new Vps proteins are involved in functions related to the prevacuole, and the network of interactions between these Class E Vps proteins will be investigated to better understand the relationship between multivesicular body formation and vesicle transport from the lateendosome/ prevacuole. This work also uncovered two new non-SNARE-like small Vps proteins predicted to span the bilayer four times, and these proteins will be investigated for a role in recruiting hydrophilic Vps protein complexes to the membranes where they function. Studies of membrane traffic in yeast have proven tremendously useful to a broader understanding of membrane transport in all eukaryotic cells because of the remarkable similarity in mechanisms and proteins that regulate these processes from yeast to humans. These basic mechanistic studies in yeast are providing important insights into our understanding of many lysosomal storage diseases in humans.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37GM032448-24
Application #
7086981
Study Section
Cell Development and Function Integrated Review Group (CDF)
Program Officer
Shapiro, Bert I
Project Start
1983-07-01
Project End
2008-06-30
Budget Start
2006-07-01
Budget End
2007-06-30
Support Year
24
Fiscal Year
2006
Total Cost
$363,746
Indirect Cost

  Institution

Name
University of Oregon
Department
Biochemistry
Type
Organized Research Units
DUNS #
948117312
City
Eugene
State
OR
Country
United States
Zip Code
97403

  Publications

Coonrod, Emily M; Graham, Laurie A; Carpp, Lindsay N et al. (2013) Homotypic vacuole fusion in yeast requires organelle acidification and not the V-ATPase membrane domain. Dev Cell 27:462-8
Coonrod, Emily M; Stevens, Tom H (2010) The yeast vps class E mutants: the beginning of the molecular genetic analysis of multivesicular body biogenesis. Mol Biol Cell 21:4057-60
Schluter, Cayetana; Lam, Karen K Y; Brumm, Jochen et al. (2008) Global analysis of yeast endosomal transport identifies the vps55/68 sorting complex. Mol Biol Cell 19:1282-94
Lottridge, Jillian M; Flannery, Andrew R; Vincelli, Jennifer L et al. (2006) Vta1p and Vps46p regulate the membrane association and ATPase activity of Vps4p at the yeast multivesicular body. Proc Natl Acad Sci U S A 103:6202-7
Bowers, Katherine; Stevens, Tom H (2005) Protein transport from the late Golgi to the vacuole in the yeast Saccharomyces cerevisiae. Biochim Biophys Acta 1744:438-54
Bowers, Katherine; Lottridge, Jillian; Helliwell, Stephen B et al. (2004) Protein-protein interactions of ESCRT complexes in the yeast Saccharomyces cerevisiae. Traffic 5:194-210
Kweon, Youngseok; Rothe, Anca; Conibear, Elizabeth et al. (2003) Ykt6p is a multifunctional yeast R-SNARE that is required for multiple membrane transport pathways to the vacuole. Mol Biol Cell 14:1868-81
Conibear, Elizabeth; Cleck, Jessica N; Stevens, Tom H (2003) Vps51p mediates the association of the GARP (Vps52/53/54) complex with the late Golgi t-SNARE Tlg1p. Mol Biol Cell 14:1610-23
Gerrard, S R; Bryant, N J; Stevens, T H (2000) VPS21 controls entry of endocytosed and biosynthetic proteins into the yeast prevacuolar compartment. Mol Biol Cell 11:613-26
Conibear, E; Stevens, T H (2000) Vps52p, Vps53p, and Vps54p form a novel multisubunit complex required for protein sorting at the yeast late Golgi. Mol Biol Cell 11:305-23

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