Journal article
Erf2, a Novel Gene Product That Affects the Localization and Palmitoylation of Ras2 in Saccharomyces cerevisiae
Molecular and cellular biology, Vol.19(10), pp.6775-6787
10/01/1999
DOI: 10.1128/MCB.19.10.6775
PMCID: PMC84674
PMID: 10490616
Abstract
Plasma membrane localization of Ras requires posttranslational addition of farnesyl and palmitoyl lipid moieties to a C-terminal C
aaX
motif (C is cysteine,
a
is any aliphatic residue,
X
is the carboxy terminal residue). To better understand the relationship between posttranslational processing and the subcellular localization of Ras, a yeast genetic screen was undertaken based on the loss of function of a palmitoylation-dependent
RAS2
allele. Mutations were identified in an uncharacterized open reading frame (
YLR246w
) that we have designated
ERF2
and a previously described suppressor of hyperactive Ras,
SHR5
.
ERF2
encodes a 41-kDa protein with four predicted transmembrane (TM) segments and a motif consisting of the amino acids Asp-His-His-Cys (DHHC) within a cysteine-rich domain (CRD), called DHHC-CRD. Mutations within the DHHC-CRD abolish Erf2 function. Subcellular fractionation and immunolocalization experiments reveal that Erf2 tagged with a triply iterated hemagglutinin epitope is an integral membrane protein that colocalizes with the yeast endoplasmic reticulum marker Kar2. Strains lacking
ERF2
are viable, but they have a synthetic growth defect in the absence of
RAS2
and partially suppress the heat shock sensitivity resulting from expression of the hyperactive
RAS2
(
V19
) allele. Ras2 proteins expressed in an
erf2
Δ strain have a reduced level of palmitoylation and are partially mislocalized to the vacuole. Based on these observations, we propose that Erf2 is a component of a previously uncharacterized Ras subcellular localization pathway. Putative members of an Erf2 family of proteins have been uncovered in yeast, plant, worm, insect, and mammalian genome databases, suggesting that Erf2 plays a role in Ras localization in all eucaryotes.
Details
- Title: Subtitle
- Erf2, a Novel Gene Product That Affects the Localization and Palmitoylation of Ras2 in Saccharomyces cerevisiae
- Creators
- Doug J. Bartels - University of IowaDavid A. Mitchell - University of IowaXiangwen Dong - University of IowaRobert J. Deschenes - University of Iowa
- Resource Type
- Journal article
- Publication Details
- Molecular and cellular biology, Vol.19(10), pp.6775-6787
- Publisher
- American Society for Microbiology
- DOI
- 10.1128/MCB.19.10.6775
- PMID
- 10490616
- PMCID
- PMC84674
- ISSN
- 0270-7306
- eISSN
- 1098-5549
- Language
- English
- Date published
- 10/01/1999
- Academic Unit
- Anatomy and Cell Biology
- Record Identifier
- 9984284342102771
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