VPS34/YLR240W Summary Help

Standard Name VPS34 1, 2
Systematic Name YLR240W
Alias END12 3 , PEP15 4 , VPL7 5 , VPT29 1 , STT8 6 , 7 , VPS7 8
Feature Type ORF, Verified
Description Phosphatidylinositol (PI) 3-kinase that synthesizes PI-3-phosphate; forms membrane-associated signal transduction complex with Vps15p to regulate protein sorting; activated by the GTP-bound form of Gpa1p; a fraction is localized, with Vps15p, to nuclear pores at nucleus-vacuole junctions and may facilitate transcription elongation for genes positioned at the nuclear periphery (9, 10, 11, 12 and see Summary Paragraph)
Name Description Vacuolar Protein Sorting 1, 2
Chromosomal Location
ChrXII:617533 to 620160 | ORF Map | GBrowse
Gbrowse
Gene Ontology Annotations All VPS34 GO evidence and references
  View Computational GO annotations for VPS34
Molecular Function
Manually curated
Biological Process
Manually curated
Cellular Component
Manually curated
High-throughput
Regulators 1 genes
Resources
Pathways
Classical genetics
conditional
dominant negative
null
reduction of function
unspecified
Large-scale survey
null
Resources
61 total interaction(s) for 47 unique genes/features.
Physical Interactions
  • Affinity Capture-MS: 9
  • Affinity Capture-RNA: 1
  • Affinity Capture-Western: 7
  • Co-fractionation: 2
  • Co-purification: 1
  • Reconstituted Complex: 1
  • Two-hybrid: 1

Genetic Interactions
  • Dosage Rescue: 1
  • Phenotypic Suppression: 1
  • Synthetic Growth Defect: 22
  • Synthetic Lethality: 15

Resources
Expression Summary
histogram
Resources
Length (a.a.) 875
Molecular Weight (Da) 100,920
Isoelectric Point (pI) 7.79
Localization
Phosphorylation PhosphoGRID | PhosphoPep Database
Structure
Homologs
sequence information
ChrXII:617533 to 620160 | ORF Map | GBrowse
SGD ORF map
Last Update Coordinates: 2011-02-03 | Sequence: 1996-07-31
Subfeature details
Relative
Coordinates
Chromosomal
Coordinates
Most Recent Updates
Coordinates Sequence
CDS 1..2628 617533..620160 2011-02-03 1996-07-31
Retrieve sequences
Analyze Sequence
S288C only
S288C vs. other species
S288C vs. other strains
Resources
External Links All Associated Seq | E.C. | Entrez Gene | Entrez RefSeq Protein | MIPS | Search all NCBI (Entrez) | UniProtKB
Primary SGDIDS000004230
SUMMARY PARAGRAPH for VPS34

VPS34 encodes a class III phosphatidylinositol (PtdIns) 3-kinase that phosphorylates phosphatidylinositol at the 3' hydroxyl position (D-3 position of the inositol ring) to produce PtdIns 3-phosphate (PtdIns 3-P) (13, 12). VPS34 was originally identified as a vacuolar protein targeting (VPT29) mutant (1), but is required for multiple protein and membrane trafficking events. These include efficient localization of a variety of vacuolar proteins (9), vacuole segregation (14), endocytosis, multivesicular body formation, constitutive autophagy via the cytoplasm-to-vacuole targeting (Cvt) pathway, and starvation-induced macroautophagy (3, 15, 16, 17, 18, 19).

Vps34p is recruited from the cytosol to the Golgi/endosome through interaction with the membrane-associated protein kinase Vps15p, which also stimulates the PtdIns 3-kinase activity of Vps34p (20, 10, 21). Interaction between Vps34p and Vps15p requires a short 28 residue-element near the C-terminus of Vps34p and two separate regions of Vps15p: the N-terminal protein kinase domain and a set of three tandem repeats of about 39 amino acids each (22). These repeat elements in Vps15p are similar to the HEAT repeats implicated in protein-protein interactions in other proteins, including Huntingtin, EF3, and Tor1p (22).

The Vps34p-Vps15p heterodimer is a component of at least two different multimeric complexes (complexes I and II), each containing Vps30p, as well as other distinct subunits that function in separate membrane trafficking processes (15). Complex I includes unique subunit Atg14p, and complex II includes Vps38p (23). Complexes I and II act in different biological processes by localizing to specific cellular locations in a manner mediated by the unique subunits of each complex (24). Complex I functions primarily in autophagy and localizes to the vacuolar membrane and the perivacuolar pre-autophagosomal structure (PAS), while complex II functions in vacuolar protein sorting and is targeted to cisternae of the late Golgi apparatus, late endosomal structures and vacuolar membranes (15, 23, 24).

Vps34p and Vps15p are also required for signalling by Gpa1p, the GTP-binding alpha subunit of the heterotrimeric G protein, at the endosome (12, 25). Gpa1p colocalizes with Vps34p and Vps15p, and binds both proteins directly in a guanine nucleotide-dependent manner (12, 25). Activated GTP-bound Gpa1p binds Vps34p, and inactive GDP-bound Gpa1p binds Vps15p (12, 25). These interactions lead to elevated production of PtdIns 3-P and promote translocation of the PtdIns 3-P binding protein Bem1p to endosomes (12, 25).

vps34 null mutants exhibit defects in protein sorting (9, 14, 26) and autophagy (15), increased sensitivity to ethanol (27, 28), and are fully defective in Gpa1p signaling, which leads to diminished efficacy and potency of mating pheromone, likely as the result of a global defect in membrane trafficking (12). Mutation of VPS34 also results in temperature-sensitive growth and defective partitioning of the vacuolar compartment between mother and daughter cells during cell division (9, 3). Cells expressing vps34-N736K do not accumulate autophagic bodies (19).

Class III phosphatidylinositol 3-kinases are conserved from yeast to human (13). The human Vps34p homolog is also found in a complex with p150, a human homolog of yeast Vps15p, which stimulates its PtdIns 3-kinase activity (3, 29). Homologs of VPS34 have been identified in other yeasts, including Candida albicans (30), Hansenula polymorpha (31) and Schizosaccharomyces pombe (32, 33). CaVps34p activity is required for C. albicans pathogenesis; C. albicans cells harboring a null allele or expressing a catalytically-inactive CaVps34 are avirulent in a mouse model for systemic candidiasis (34).

About Phosphatidylinositol Phosphate Biosynthesis

The phosphorylated products of phosphatidylinositol (PtdIns, PI), collectively referred to as phosphoinositides or phosphatidylinositol phosphates (PtdInsPs, PIPs), are membrane-bound lipids that function as structural components of membranes, as well as regulators of many cellular processes in eukaryotes, including vesicle-mediated membrane trafficking, cell wall integrity, and actin cytoskeleton organization (reviewed in 18 and 35). PtdInsPs are also precursors of the water-soluble inositol phosphates (IPs), an important class of intracellular signaling molecules (reviewed in 36, 37 and 38).

The inositol ring of the membrane phospholipids and the water-soluble IPs are readily phosphorylated and dephosphorylated at a number of positions making them well suited as key regulators. PtdIns can be phosphorylated at one or a combination of positions (3', 4', or 5') on the inositol headgroup, generating a set of unique stereoisomers that have specific biological functions (reviewed in 18). These stereoisomers have been shown to be restricted to certain membranes (reviewed in 18). Phosphatidylinositol 4-phosphate (PtdIns4P) is the major PtdInsP species of the Golgi apparatus, where it plays a role in the vesicular trafficking of secretory proteins from the Golgi to the plasma membrane (reviewed in 18). Phosphatidylinositol 4,5-bisphosphate (PtdIns[4,5]P2) is the major species found at the plasma membrane and is involved in the regulation of actin cytoskeleton organization, as well as cell wall integrity, and heat shock response pathways (reviewed in 18). Phosphatidylinositol 3-phosphate (PtdIns3P) is found predominantly at endosomal membranes and in multivesicular bodies (MVB), where it plays a role in endosomal and vacuolar membrane trafficking. Phosphatidylinositol 3,5-bisphosphate (PtdIns[3,5]P2) is found on vacuolar membranes where it plays an important role in the MVB sorting pathway (reviewed in 18).

Phosphorylation and dephosphorylation of the inositol headgroups of PtdInsPs at specific membrane locations signals the recruitment of certain proteins essential for vesicular transport (35, and reviewed in 18). PtdInsPs recruit proteins that contain PtdInsP-specific binding domains, such as the well-studied pleckstrin homology (PH) domain that recognizes the phosphorylation pattern of specific PtdInsP inositol headgroups (reviewed in 18).

A number of kinases and phosphatases are involved in the generation and interconversions of PtdInsPs, the majority of which have been well conserved during evolution (reviewed in 18). The PtdInsP kinases, in contrast to the lipid phosphatases, have a higher degree of specificity. While each kinase appears to phosphorylate only one substrate, many of the lipid phosphatases can dephosphorylate a number of substrates.

Last updated: 2008-05-08 Contact SGD

References cited on this page View Complete Literature Guide for VPS34
1) Robinson JS, et al.  (1988) Protein sorting in Saccharomyces cerevisiae: isolation of mutants defective in the delivery and processing of multiple vacuolar hydrolases. Mol Cell Biol 8(11):4936-48
2) Rothman JH, et al.  (1989) Characterization of genes required for protein sorting and vacuolar function in the yeast Saccharomyces cerevisiae. EMBO J 8(7):2057-65
3) Munn AL and Riezman H  (1994) Endocytosis is required for the growth of vacuolar H(+)-ATPase-defective yeast: identification of six new END genes. J Cell Biol 127(2):373-86
4) Stepp JD, et al.  (1997) The yeast adaptor protein complex, AP-3, is essential for the efficient delivery of alkaline phosphatase by the alternate pathway to the vacuole. J Cell Biol 139(7):1761-74
5) Rothman JH and Stevens TH  (1986) Protein sorting in yeast: mutants defective in vacuole biogenesis mislocalize vacuolar proteins into the late secretory pathway. Cell 47(6):1041-51
6) Yoshida S, et al.  (1992) Characterization of a staurosporine- and temperature-sensitive mutant, stt1, of Saccharomyces cerevisiae: STT1 is allelic to PKC1. Mol Gen Genet 231(3):337-44
7) Yoshida S and Anraku Y  (2000) Characterization of staurosporine-sensitive mutants of Saccharomyces cerevisiae: vacuolar functions affect staurosporine sensitivity. Mol Gen Genet 263(5):877-88
8) Raymond CK, et al.  (1992) Morphological classification of the yeast vacuolar protein sorting mutants: evidence for a prevacuolar compartment in class E vps mutants. Mol Biol Cell 3(12):1389-402
9) Herman PK and Emr SD  (1990) Characterization of VPS34, a gene required for vacuolar protein sorting and vacuole segregation in Saccharomyces cerevisiae. Mol Cell Biol 10(12):6742-54
10) Stack JH, et al.  (1993) A membrane-associated complex containing the Vps15 protein kinase and the Vps34 PI 3-kinase is essential for protein sorting to the yeast lysosome-like vacuole. EMBO J 12(5):2195-204
11) Burda P, et al.  (2002) Retromer function in endosome-to-Golgi retrograde transport is regulated by the yeast Vps34 PtdIns 3-kinase. J Cell Sci 115(Pt 20):3889-900
12) Slessareva JE, et al.  (2006) Activation of the phosphatidylinositol 3-kinase Vps34 by a G protein alpha subunit at the endosome. Cell 126(1):191-203
13) Auger KR, et al.  (1989) Phosphatidylinositol 3-kinase and its novel product, phosphatidylinositol 3-phosphate, are present in Saccharomyces cerevisiae. J Biol Chem 264(34):20181-4
14) Schu PV, et al.  (1993) Phosphatidylinositol 3-kinase encoded by yeast VPS34 gene essential for protein sorting. Science 260(5104):88-91
15) Kihara A, et al.  (2001) Two distinct Vps34 phosphatidylinositol 3-kinase complexes function in autophagy and carboxypeptidase Y sorting in Saccharomyces cerevisiae. J Cell Biol 152(3):519-30
16) Wurmser AE and Emr SD  (2002) Novel PtdIns(3)P-binding protein Etf1 functions as an effector of the Vps34 PtdIns 3-kinase in autophagy. J Cell Biol 158(4):761-72
17) Katzmann DJ, et al.  (2003) Vps27 recruits ESCRT machinery to endosomes during MVB sorting. J Cell Biol 162(3):413-23
18) Strahl T and Thorner J  (2007) Synthesis and function of membrane phosphoinositides in budding yeast, Saccharomyces cerevisiae. Biochim Biophys Acta 1771(3):353-404
19) Obara K, et al.  (2008) Transport of phosphatidylinositol 3-phosphate into the vacuole via autophagic membranes in Saccharomyces cerevisiae. Genes Cells 13(6):537-47
20) Herman PK, et al.  (1991) A novel protein kinase homolog essential for protein sorting to the yeast lysosome-like vacuole. Cell 64(2):425-37
21) Stack JH, et al.  (1995) Vesicle-mediated protein transport: regulatory interactions between the Vps15 protein kinase and the Vps34 PtdIns 3-kinase essential for protein sorting to the vacuole in yeast. J Cell Biol 129(2):321-34
22) Budovskaya YV, et al.  (2002) The C terminus of the Vps34p phosphoinositide 3-kinase is necessary and sufficient for the interaction with the Vps15p protein kinase. J Biol Chem 277(1):287-94
23) Klionsky DJ  (2005) The molecular machinery of autophagy: unanswered questions. J Cell Sci 118(Pt 1):7-18
24) Obara K, et al.  (2006) Assortment of phosphatidylinositol 3-kinase complexes--Atg14p directs association of complex I to the pre-autophagosomal structure in Saccharomyces cerevisiae. Mol Biol Cell 17(4):1527-39
25) Slessareva JE and Dohlman HG  (2006) G protein signaling in yeast: new components, new connections, new compartments. Science 314(5804):1412-3
26) Deloche O, et al.  (2001) Vps10p transport from the trans-Golgi network to the endosome is mediated by clathrin-coated vesicles. Mol Biol Cell 12(2):475-85
27) Takahashi T, et al.  (2001) Identification of genes required for growth under ethanol stress using transposon mutagenesis in Saccharomyces cerevisiae. Mol Genet Genomics 265(6):1112-9
28) van Voorst F, et al.  (2006) Genome-wide identification of genes required for growth of Saccharomyces cerevisiae under ethanol stress. Yeast 23(5):351-9
29) Panaretou C, et al.  (1997) Characterization of p150, an adaptor protein for the human phosphatidylinositol (PtdIns) 3-kinase. Substrate presentation by phosphatidylinositol transfer protein to the p150.Ptdins 3-kinase complex. J Biol Chem 272(4):2477-85
30) Eck R, et al.  (2000) A phosphatidylinositol 3-kinase of Candida albicans: molecular cloning and characterization. Yeast 16(10):933-44
31) Kiel JA, et al.  (1999) The Hansenula polymorpha PDD1 gene product, essential for the selective degradation of peroxisomes, is a homologue of Saccharomyces cerevisiae Vps34p. Yeast 15(9):741-54
32) Kimura K, et al.  (1995) Phosphatidylinositol-3 kinase in fission yeast: a possible role in stress responses. Biosci Biotechnol Biochem 59(4):678-82
33) Takegawa K, et al.  (1995) Schizosaccharomyces pombe Vps34p, a phosphatidylinositol-specific PI 3-kinase essential for normal cell growth and vacuole morphology. J Cell Sci 108 ( Pt 12)():3745-56
34) Gunther J, et al.  (2005) Generation and functional in vivo characterization of a lipid kinase defective phosphatidylinositol 3-kinase Vps34p of Candida albicans. Microbiology 151(Pt 1):81-9
35) De Camilli P, et al.  (1996) Phosphoinositides as regulators in membrane traffic. Science 271(5255):1533-9
36) York JD  (2006) Regulation of nuclear processes by inositol polyphosphates. Biochim Biophys Acta 1761(5-6):552-9
37) Bennett M, et al.  (2006) Inositol pyrophosphates: metabolism and signaling. Cell Mol Life Sci 63(5):552-64
38) Bhandari R, et al.  (2007) Inositol pyrophosphate pyrotechnics. Cell Metab 5(5):321-3