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Formation of She2p tetramers is required for mRNA binding, mRNP assembly, and localization.
RNA 15, 2002-2012 (2009)
In eukaryotic cells, dozens to hundreds of different mRNAs are localized by specialized motor-dependent transport complexes. One of the best-studied examples for directional mRNA transport is the localization of ASH1 mRNA in Saccharomyces cerevisiae. For transport, ASH1 mRNA is bound by the unusual RNA-binding protein She2p. Although previous results indicated that She2p forms dimers required for RNA binding and transcript localization, it remained unclear if the dimer constitutes the minimal RNA-binding unit assembling in vivo. By using analytical ultracentrifugation we found that She2p forms larger oligomeric complexes in solution. We also identified a point mutant that shows impaired oligomer formation. Size-exclusion chromatography suggests that She2p forms defined tetramers at physiological concentrations. Subsequent structural studies by small-angle X-ray scattering confirmed this finding and demonstrated that the previously observed She2p dimers interact in a head-to-head conformation to form an elongated tetrameric complex. This She2p tetramer suggests the generation of large continuous RNA-binding surfaces at both sides of the complex. Biochemical studies and immunostaining of cells confirmed that She2p tetramer formation is required for RNA binding, efficient mRNP assembly, and mRNA localization in vivo. Our finding on She2p tetramerization resolves previously raised questions on complex formation and mRNP function.
Impact Factor
Scopus SNIP
Web of Science
Times Cited
Times Cited
Scopus
Cited By
Cited By
Altmetric
5.018
1.500
29
35
Anmerkungen
Besondere Publikation
Auf Hompepage verbergern
Publikationstyp
Artikel: Journalartikel
Dokumenttyp
Wissenschaftlicher Artikel
Schlagwörter
She2p; She3p; mRNP; RNA localization; RNA binding; SAXS
Sprache
englisch
Veröffentlichungsjahr
2009
HGF-Berichtsjahr
2009
ISSN (print) / ISBN
1355-8382
e-ISSN
1469-9001
Zeitschrift
RNA
Quellenangaben
Band: 15,
Heft: 11,
Seiten: 2002-2012
Verlag
Cold Spring Harbor Laboratory Press
Verlagsort
U.S.
Begutachtungsstatus
Peer reviewed
PSP-Element(e)
G-551400-001
G-501700-003
G-501700-003
PubMed ID
19710186
Scopus ID
70350120574
Erfassungsdatum
2009-12-31