Arf1p provides an unexpected link between COPI vesicles and mRNA in Saccharomyces cerevisiae.
The small GTPase Arf1p is involved in different cellular processes that require its accumulation at specific cellular locations. The recruitment of Arf1p to distinct points of action might be achieved by association of Arf1p with different proteins. To identify new interactors of Arf1p, we performed an affinity chromatography with GTP- or GDP-bound Arf1p proteins. A new interactor of Arf1p-GTP was identified as Pab1p, which binds to the polyA-tail of mRNAs. Pab1p was found to associate with purified COPI-coated vesicles generated from Golgi membranes in vitro. The stability of the Pab1p-Arf1p complex depends on the presence of mRNA. Both symmetrically distributed mRNAs as well as the asymmetrically localized ASH1 mRNA are found in association with Arf1p. Remarkably, Arf1p and Pab1p are both required to restrict ASH1 mRNA to the bud tip. Arf1p and coatomer play an unexpected role in localizing mRNA independent and downstream of the SHE machinery. Hereby acts the SHE machinery in long-range mRNA transport, whereas COPI vesicles could act as short-range and localization vehicles. The endoplasmic reticulum (ER)-Golgi shuttle might be involved in concentrating mRNA at the ER.
Trautwein
Mark
M
Friedrich Miescher Laboratorium, Max Planck Gesellschaft, D-72076 Tübingen, Germany.
Dengjel
Jörn
J
Schirle
Markus
M
Spang
Anne
A
D020823:ADP-Ribosylation Factor 1 / Q000737:chemistry / Q000502:physiology*; D000199:Actins / Q000378:metabolism; D001692:Biological Transport; D015152:Blotting, Northern; D002846:Chromatography, Affinity; D020755:Coat Protein Complex I / Q000378:metabolism*; D003599:Cytoskeleton / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D005838:Genotype; D006056:Golgi Apparatus / Q000378:metabolism; D008954:Models, Biological; D009154:Mutation; D011003:Ploidies; D011061:Poly A / Q000737:chemistry; D039102:Poly(A)-Binding Proteins / Q000378:metabolism; D011485:Protein Binding; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D013696:Temperature; D014158:Transcription, Genetic; D020798:Two-Hybrid System Techniques
D000199:Actins; D020755:Coat Protein Complex I; C083327:PUB1 protein, S cerevisiae; D039102:Poly(A)-Binding Proteins; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins; D011061:Poly A; D020823:ADP-Ribosylation Factor 1