Conservation of Structure and Subunit Interactions in Yeast Homologues of Splicing Factor 3b (SF3b) Subunits

Overview

Journal RNA

Specialty Molecular Biology

Date 1998 Jan 22

PMID 9436903

Citations 33

Authors

H IGEL

S Wells

R Perriman

M Ares Jr

Affiliations

Soon will be listed here.

Abstract

Human SAP 49, a subunit of the multimeric splicing factor 3b (SF3b), contains two RNA recognition motifs (RRMs) and binds another SF3b subunit called SAP 145, whose yeast homologue is CUS1. Here we show that the predicted yeast open reading frame YOR319w (HSH49) encodes an essential yeast splicing factor. Using bacterially expressed proteins, we find that yeast HSH49 binds CUS1. Mutations that alter putative RNA-binding residues of either HSH49 RRM are lethal in vivo, but do not prevent binding to CUS1 in vitro, suggesting that the predicted RNA-binding surfaces of HSH49 are not required for interaction with CUS1. In vivo interaction tests show that HSH49 and CUS1 associate primarily through the N-terminal RRM of HSH49. Recombinant HSH49 protein has a general RNA-binding activity that does not require CUS1. The parallels in structure and interaction between two SF3b subunits from yeast implies that the mechanism of SF3b action is highly conserved.

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References

Hodges P, Beggs J . RNA splicing. U2 fulfils a commitment. Curr Biol. 1994; 4(3):264-7. DOI: 10.1016/s0960-9822(00)00061-0. View

Rain J, Legrain P . Toward a functional analysis of the yeast genome through exhaustive two-hybrid screens. Nat Genet. 1997; 16(3):277-82. DOI: 10.1038/ng0797-277. View

Oubridge C, Ito N, Evans P, Teo C, Nagai K . Crystal structure at 1.92 A resolution of the RNA-binding domain of the U1A spliceosomal protein complexed with an RNA hairpin. Nature. 1994; 372(6505):432-8. DOI: 10.1038/372432a0. View

Madhani H, Guthrie C . Dynamic RNA-RNA interactions in the spliceosome. Annu Rev Genet. 1994; 28:1-26. DOI: 10.1146/annurev.ge.28.120194.000245. View

Ares Jr M, Weiser B . Rearrangement of snRNA structure during assembly and function of the spliceosome. Prog Nucleic Acid Res Mol Biol. 1995; 50:131-59. DOI: 10.1016/s0079-6603(08)60813-2. View

Nagai K, Oubridge C, Ito N, Avis J, Evans P . The RNP domain: a sequence-specific RNA-binding domain involved in processing and transport of RNA. Trends Biochem Sci. 1995; 20(6):235-40. DOI: 10.1016/s0968-0004(00)89024-6. View

Sayle R . RASMOL: biomolecular graphics for all. Trends Biochem Sci. 1995; 20(9):374. DOI: 10.1016/s0968-0004(00)89080-5. View

Wells S, Neville M, Haynes M, Wang J, IGEL H, Ares Jr M . CUS1, a suppressor of cold-sensitive U2 snRNA mutations, is a novel yeast splicing factor homologous to human SAP 145. Genes Dev. 1996; 10(2):220-32. DOI: 10.1101/gad.10.2.220. View

Gozani O, Feld R, REED R . Evidence that sequence-independent binding of highly conserved U2 snRNP proteins upstream of the branch site is required for assembly of spliceosomal complex A. Genes Dev. 1996; 10(2):233-43. DOI: 10.1101/gad.10.2.233. View

10.

VanHoy R, Wise J . Molecular analysis of a novel schizosaccharomyces pombe gene containing two RNP consensus-sequence RNA-binding domains. Curr Genet. 1996; 29(4):307-15. DOI: 10.1007/BF02208611. View

11.

Yan D, Ares Jr M . Invariant U2 RNA sequences bordering the branchpoint recognition region are essential for interaction with yeast SF3a and SF3b subunits. Mol Cell Biol. 1996; 16(3):818-28. PMC: 231062. DOI: 10.1128/MCB.16.3.818. View

12.

Higgins D, Thompson J, Gibson T . Using CLUSTAL for multiple sequence alignments. Methods Enzymol. 1996; 266:383-402. DOI: 10.1016/s0076-6879(96)66024-8. View

13.

Henry M, Borland C, Bossie M, Silver P . Potential RNA binding proteins in Saccharomyces cerevisiae identified as suppressors of temperature-sensitive mutations in NPL3. Genetics. 1996; 142(1):103-15. PMC: 1206938. DOI: 10.1093/genetics/142.1.103. View

14.

Kramer A . The structure and function of proteins involved in mammalian pre-mRNA splicing. Annu Rev Biochem. 1996; 65:367-409. DOI: 10.1146/annurev.bi.65.070196.002055. View

15.

Tanaka Y, Ohta A, Terashima K, Sakamoto H . Polycistronic expression and RNA-binding specificity of the C. elegans homologue of the spliceosome-associated protein SAP49. J Biochem. 1997; 121(4):739-45. DOI: 10.1093/oxfordjournals.jbchem.a021648. View

16.

Ruiz-Lozano P, Doevendans P, Brown A, Gruber P, Chien K . Developmental expression of the murine spliceosome-associated protein mSAP49. Dev Dyn. 1997; 208(4):482-90. DOI: 10.1002/(SICI)1097-0177(199704)208:4<482::AID-AJA4>3.0.CO;2-E. View

17.

Bradford M . A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72:248-54. DOI: 10.1016/0003-2697(76)90527-3. View

18.

Milligan J, Uhlenbeck O . Synthesis of small RNAs using T7 RNA polymerase. Methods Enzymol. 1989; 180:51-62. DOI: 10.1016/0076-6879(89)80091-6. View

19.

Altschul S, Gish W, Miller W, Myers E, Lipman D . Basic local alignment search tool. J Mol Biol. 1990; 215(3):403-10. DOI: 10.1016/S0022-2836(05)80360-2. View

20.

Kolodziej P, Young R . Epitope tagging and protein surveillance. Methods Enzymol. 1991; 194:508-19. DOI: 10.1016/0076-6879(91)94038-e. View