1. The Yeast hnRNP-Like Proteins Yra1p and Yra2p Participate in mRNA Export through Interaction with Mex67p
- Author
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Patrizia Vinciguerra, Daniel Zenklusen, Françoise Stutz, and Yvan Strahm
- Subjects
Nucleocytoplasmic Transport Proteins ,Saccharomyces cerevisiae Proteins ,RNA, Messenger/genetics/metabolism ,Recombinant Fusion Proteins ,Saccharomyces cerevisiae ,Immunoblotting ,Plasma protein binding ,Fungal Proteins/chemistry/genetics/metabolism ,Nuclear Proteins/chemistry/genetics/metabolism ,DNA-binding protein ,Recombinant Fusion Proteins/genetics/metabolism ,Fungal Proteins ,Transformation, Genetic ,Genes, Reporter ,Genes, Reporter/genetics ,Nucleocytoplasmic Communication ,RNA, Messenger ,RNA-Binding Proteins/genetics/metabolism ,Nuclear protein ,Molecular Biology ,Ribonucleoproteins/chemistry/genetics/metabolism ,Genetics ,Messenger RNA ,biology ,RNA ,Nuclear Proteins ,RNA-Binding Proteins ,RNA, Fungal ,Cell Biology ,biology.organism_classification ,Plasmids/genetics/metabolism ,In vitro ,Cell biology ,Protein Structure, Tertiary ,Ribonucleoproteins ,Saccharomyces cerevisiae/genetics/metabolism ,RNA, Fungal/metabolism ,Plasmids ,Protein Binding - Abstract
Yra1p is an essential nuclear protein which belongs to the evolutionarily conserved REF (RNA and export factor binding proteins) family of hnRNP-like proteins. Yra1p contributes to mRNA export in vivo and directly interacts with RNA and the shuttling mRNP export receptor Mex67p in vitro. Here we describe a second nonessential Saccharomyces cerevisiae family member, called Yra2p, which is able to complement a YRA1 deletion when overexpressed. Like other REF proteins, Yra1p and Yra2p consist of two highly conserved N- and C-terminal boxes and a central RNP-like RNA-binding domain (RBD). These conserved regions are separated by two more variable regions, N-vr and C-vr. Surprisingly, the deletion of a single conserved box or the deletion of the RBD in Yra1p does not affect viability. Consistently, neither the conserved N and C boxes nor the RBD is required for Mex67p and RNA binding in vitro. Instead, the N-vr and C-vr regions both interact with Mex67p and RNA. We further show that Yra1 deletion mutants which poorly interact with Mex67p in vitro affect the association of Mex67p with mRNP complexes in vivo and are paralleled by poly(A)(+) RNA export defects. These observations support the idea that Yra1p promotes mRNA export by facilitating the recruitment of Mex67p to the mRNP.
- Published
- 2001