Identification of a yeast snRNP protein and detection of snRNP-snRNP interactions

Lossky, M; Anderson, Gordon J.; Jackson, Stephen; Beggs, Jean D.

doi:10.1016/0092-8674(87)90588-5

Cited by 182 publications

(184 citation statements)

References 38 publications

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“…No free U6 snRNP was found and very little amounts of U4/U6 snRNP and [U4/U6.U5] tri-snRNP were detected in this extract (compare fractions 6-8, 12-14 and 24-28 in Figures 7C and D, respectively). In contrast, the free U5 snRNP amount (fractions [16][17][18][19][20][21][22] was increased and a free U4 snRNP was found (fractions [8][9][10][11][12][13][14]. This particle has a reduced velocity compared to the U4/U6 snRNP and very probably derives from U4/U6 snRNP disruption.…”

Section: Immunodetectionmentioning

confidence: 85%

“…In mammalian cells, about ten proteins have been found associated with the U4/U6 snRNP, none of which, however, is specific to this particle (13). In addition, several proteins have been shown to participate to the formation of the [U4/U6.U5] tri-snRNP, including the U5 snRNP specific protein, homologous to the yeast PRP8 protein (14)(15)(16)(17)(18)(19).…”

Section: Introductionmentioning

confidence: 99%

“…However, genetic approaches have led to the isolation of many splicing deficient mutants, called prp mutants (20,21). Some PRP genes encode snRNP components, such as PRP4, PRP8 and PRP24 (14,19,(22)(23)(24) and others, such as PRP2, PRP5, PRP9, PRP11, PRP16, PRP22 and PRP28, are implicated in spliceosome assembly or disassembly (25)(26)(27)(28)(29)(30)(31).…”

Section: Introductionmentioning

confidence: 99%

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The biochemical defects ofprp4-1andprp6-1yeast splicing mutants reveal that the PRP6 protein is required for the accumulation of the [U4/U6.U5] tri-snRNP

Galisson

Legrain

1993

Nucl Acids Res

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Section: Immunodetectionmentioning

confidence: 85%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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The biochemical defects ofprp4-1andprp6-1yeast splicing mutants reveal that the PRP6 protein is required for the accumulation of the [U4/U6.U5] tri-snRNP

Galisson

Legrain

1993

Nucl Acids Res

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“…Yeast strains, plasmids and RNA substrates S.cerevisiae strains BJ2412 and SPJ8.31 were described previously (12,17) and DJY85 (MATa/%cx prp2-1/prp2-1, ura3/ura3, adel/ADEJ, ade2/ade2, trpJfJRPI, his3/HIS3, tyri/TYRI, lys2-801/LYS2-801, canJICANI) was obtained from D. J. Jamieson, Dundee.…”

Section: Methodsmentioning

confidence: 99%

Interaction of the yeast splicing factor PRP8 with substrate RNA during both steps of splicing

1995

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PRP8 protein of Saccharomyces cerevisiae interacts directly with pre-mRNA in spliceosomes, shown previously by UV-crosslinking. To analyse at which steps of splicing and with which precursor-derived RNA species the interaction(s) take place, UV-crosslinking was combined with PRP8-specific immunoprecipitation and the coprecipitated RNA species were analysed. Specific precipitation of intron-exon 2 and excised intron species was observed. PRP8 protein could be UV-crosslinked to pre-mRNA in PRP2-depleted spliceosomes stalled before initiation of the splicing reaction. Thus, the interaction of PRP8 protein with substrate RNA is established prior to the first transesterification reaction, is maintained during both steps of splicing and continues with the excised intron after completion of the splicing reaction. RNase Ti treatment of spliceosomes revealed that substrate RNA fragments of the 5' splice site region and the branchpoint-3' splice site region could be coimmunoprecipitated with PRP8 specific antibodies, indicating that these are potential sites of interaction for PRP8 protein with substrate RNA. Protection of the branchpoint-3' splice site region was detected only after step 1 of splicing. The results allow a first glimpse at the pattem of PRP8 protein-RNA interactions during splicing and provide a fundamental basis for future analysis of these interactions.

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“…Using immunological tools, Last and Woolford (1986) have shown that the protein products of the RNA 2 and RNA 3 genes are nuclear proteins of -100,000 and 55,000 daltons, respectively. The RNA 8 gene specifies a 260,000-dalton protein that is complexed to U5 snRNA and, after incubation in the pres ence of ATP, also to U4 and U6 snRNAs (Lossky et al 1987).…”

mentioning

confidence: 99%

Identification and purification of a 62,000-dalton protein that binds specifically to the polypyrimidine tract of introns.

García-Blanco

Jamison

Sharp

1989

Genes & Development

345

296

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A protein of molecular size 62,000 daltons (p62) was detected in HeLa cell nuclear extracts by UV cross-linking to mRNA precursors. p62 binds specifically to the polypyrimidine tract of the 3' splice site region of introns. p62 purified to homogeneity binds the polypyrimidine tract of pre-mRNAs. This binding does not require the AG dinucleotide at the 3' splice site. Alterations in the polypyrimidine tract that reduce the binding of p62 yield a corresponding reduction in the efficiency of formation of a U2 snRNP/pre-mRNA complex and splicing. The p62 protein is retained in the spliceosome, where it remains bound to the pre-mRNA. This polypyrimidine tract binding protein (pPTB) is proposed to be a critical component in recognition of the 3' splice site during splicing.

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Identification of a yeast snRNP protein and detection of snRNP-snRNP interactions

Cited by 182 publications

References 38 publications

The biochemical defects ofprp4-1andprp6-1yeast splicing mutants reveal that the PRP6 protein is required for the accumulation of the [U4/U6.U5] tri-snRNP

The biochemical defects ofprp4-1andprp6-1yeast splicing mutants reveal that the PRP6 protein is required for the accumulation of the [U4/U6.U5] tri-snRNP

Interaction of the yeast splicing factor PRP8 with substrate RNA during both steps of splicing

Identification and purification of a 62,000-dalton protein that binds specifically to the polypyrimidine tract of introns.

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