Functional Cus1p is found with Hsh155p in a multiprotein splicing factor associated with U2 snRNA.
To explore the dynamics of snRNP structure and function, we have studied Cus1p, identified as a suppressor of U2 snRNA mutations in budding yeast. Cus1p is homologous to human SAP145, a protein present in the 17S form of the human U2 snRNP. Here, we define the Cus1p amino acids required for function in yeast. The segment of Cus1p required for binding to Hsh49p, a homolog of human SAP49, is contained within an essential region of Cus1p. Antibodies against Cus1p coimmunoprecipitate U2 snRNA, as well as Hsh155p, a protein homologous to human SAP155. Biochemical fractionation of splicing extracts and reconstitution of heat-inactivated splicing extracts from strains carrying a temperature-sensitive allele of CUS1 indicate that Cus1p and Hsh155p reside in a functional, high-salt-stable complex that is salt-dissociable from U2 snRNA. We propose that Cus1p, Hsh49p, and Hsh155p exist in a stable protein complex which can exchange with a core U2 snRNP and which is necessary for U2 snRNP function in prespliceosome assembly. The Cus1p complex shares functional as well as structural similarities with human SF3b.
Pauling
M H
MH
Department of Biology, Center for Molecular Biology of RNA, Sinsheimer Laboratories, University of California, Santa Cruz, Santa Cruz, California 95064, USA.
McPheeters
D S
DS
Ares
M
M
D048868:Adaptor Proteins, Signal Transducing; D000595:Amino Acid Sequence; D005656:Fungal Proteins / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D010750:Phosphoproteins / Q000235:genetics; D012326:RNA Splicing; D000072260:RNA Splicing Factors; D012342:RNA, Small Nuclear / Q000235:genetics*; D016601:RNA-Binding Proteins; D017413:Ribonucleoprotein, U2 Small Nuclear / Q000235:genetics; D012269:Ribosomal Proteins / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins; D016415:Sequence Alignment
D048868:Adaptor Proteins, Signal Transducing; C097976:CUS1 protein, S cerevisiae; D005656:Fungal Proteins; C405527:HSH155 protein, S cerevisiae; D010750:Phosphoproteins; D000072260:RNA Splicing Factors; D012342:RNA, Small Nuclear; D016601:RNA-Binding Proteins; D017413:Ribonucleoprotein, U2 Small Nuclear; D012269:Ribosomal Proteins; C491800:SAP155 protein, S cerevisiae; C112491:SF3B1 protein, human; D029701:Saccharomyces cerevisiae Proteins