Phosphorylation of spliceosomal protein SAP 155 coupled with splicing catalysis
Genes & DevelopmentPublished 15 May 1998Open access
Changyu Wang, Katrin F. Chua, Wolfgang Seghezzi, Emma Lees, Or Gozani, Robin Reed
Citations195
SJR quartileQ1
SJR score4.00
SNIP1.43
Generate an AI Snapshot to get a quick, structured summary of this paper.
Study Snapshot
ObjectiveStudy objective
MethodsResearch methodology
PopulationPopulation studied
Sample sizeSample sizes
OutcomesStudy outcomes here
ResultsStudy results comes here
LimitationsResearch study limitations comes here
A concise AI-generated summary of the paper will appear here once you click Generate AI Snapshot.
TL;DR
The carboxy-terminal two-thirds of SAP 155 shows the highest conservation and is remarkably similar to the regulatory subunit A of the phosphatase PP2A, making this the first example of a protein modification tightly regulated with splicing catalysis.
Abstract
The U2 snRNP component SAP 155 contacts pre-mRNA on both sides of the branch site early in spliceosome assembly and is therefore positioned near or at the spliceosome catalytic center. We have isolated a cDNA encoding human SAP 155 and identified its highly related Saccharomyces cerevisiae homolog (50% identity). The carboxy-terminal two-thirds of SAP 155 shows the highest conservation and is remarkably similar to the regulatory subunit A of the phosphatase PP2A. Significantly, SAP 155 is phosphorylated concomitant with or just after catalytic step one, making this the first example of a protein modification tightly regulated with splicing catalysis.
Keywords
Biochemistry, Genetics and Molecular Biology
Annual Review of BiochemistryTHE STRUCTURE AND FUNCTION OF PROTEINS INVOLVED IN MAMMALIAN PRE-mRNA SPLICING
689 Citations1996Angela Krämer
The EMBO JournalThe Clk/Sty protein kinase phosphorylates SR splicing factors and regulates their intranuclear distribution.
575 Citations1996Karen Colwill, Tony Pawson +5 more
Results suggest that Clk/Sty kinase directly regulates the activity and compartmentalization of SR splicing factors.
ScienceSite-Specific Modification of Pre-mRNA: the 2′-Hydroxyl Groups at the Splice Sites
469 Citations1992Melissa J. Moore, Phillip A. Sharp
Splicing of the modified pre-messenger RNA's in vitro revealed that, although a 2'-hydroxyl is not absolutely required at either splice site, the 2'-Hydroxyl at the 3' splicing site is important for the second step of splicing.
Genes & DevelopmentPhosphorylation of the ASF/SF2 RS domain affects both protein-protein and protein-RNA interactions and is necessary for splicing.
402 Citations1997Shou-Hua Xiao, James L. Manley
Evidence is provided that the RS domain of unphosphorylated recombinant ASF/SF2 is necessary, but not sufficient, for binding to the U1 snRNP-specific 70-kD protein (70K) in vitro, and conditions that reveal for the first time a phosphorylation requirement for ASF /SF2 splicing activity in vitro.
The Journal of Cell BiologyA conserved family of nuclear phosphoproteins localized to sites of polymerase II transcription.
340 Citations1991Michael Roth, Alan M. Zahler +1 more
An antibody was identified previously that recognizes sites of polymerase II transcription on lampbrush chromosomes, puffs on polytene chromosomes, and many small granules in the nucleoplasm of all cells tested, which suggests that this protein plays a role in pre-mRNA splicing.
The EMBO JournalRegulation of mammalian spliceosome assembly by a protein phosphorylation mechanism.
331 Citations1994Jacqueline E. Mermoud, Paul Cohen +1 more
It appears that pre‐mRNA splicing, in common with other biological processes, can be regulated both positively and negatively by reversible protein phosphorylation.
Current Opinion in Genetics & DevelopmentInitial splice-site recognition and pairing during pre-mRNA splicing
281 Citations1996Robin Reed
Highlights include the demonstration that exonic enhancers can promote trans splicing and that an excess of the serine and arginine rich family of splicing proteins can obviate the requirement for U1 small nuclear ribonucleoprotein particle in splicing.
Current Opinion in Cell BiologyProtein functions in pre-mRNA splicing
270 Citations1997Cindy L. Will, Reinhard Lührmann
Growing evidence suggests that proteins may contribute directly to the spliceosome's active sites.
Genes & DevelopmentEvidence that sequence-independent binding of highly conserved U2 snRNP proteins upstream of the branch site is required for assembly of spliceosomal complex A.
254 Citations1996Or Gozani, R Feld +1 more
Analysis of proteins that cross-link to 32P-site-specifically labeled pre-mRNA in the A complex indicates sequence-independent binding of the highly conserved SF3a/SF3b subunits upstream of the branch site is essential for anchoring U2 snRNP to pre- mRNA.
The EMBO JournalA novel set of spliceosome‐associated proteins and the essential splicing factor PSF bind stably to pre‐mRNA prior to catalytic step II of the splicing reaction.
220 Citations1994Or Gozani, James G. Patton +1 more
It is shown that a novel set of 14 spliceosome‐associated proteins (SAPs) and the essential splicing factor PSF are specifically associated with the C complex, implicating these proteins in catalytic step II.
Nucleic Acids ResearchSer/Thr-specific protein phosphatases are required for both catalytic steps of pre-mRNA splicing
217 Citations1992Jacqueline E. Mermoud, Philip Cohen +1 more
Protein phosphatase activity was not required for efficient assembly of splicing complexes containing each of the U1, U2, U4/U6 and U5 snRNPs, indicating that reversible protein phosphorylation may play an important role in regulating the pre-mRNA splicing mechanism.
Proceedings of the National Academy of SciencesPurification and characterization of a kinase specific for the serine- and arginine-rich pre-mRNA splicing factors.
203 Citations1994Jian‐Fang Gui, Hélène Tronchère +2 more
It is shown that SR PK1 restores both mobility and mAB 104 reactivity to a SR protein SF2/ASF (splicing factor 2/alternative splicing factor) produced in bacteria, suggesting that SRPK1 is responsible for the generation of the mAb 104-specific phosphoepitope in vivo.
Genes & DevelopmentThe yeast MUD2 protein: an interaction with PRP11 defines a bridge between commitment complexes and U2 snRNP addition.
186 Citations1994Nadja Abovich, X C Liao +1 more
The experiments identify a specific inter-snRNP protein-protein contact that occurs during spliceosome assembly and more generally support substantial functional similarity between U2AF65 and MUD2P.
NatureThiophosphorylation of U1-70K protein inhibits pre-mRNA splicing
150 Citations1993Jamal Tazi, Ute Kornstädt +5 more
The data show that the state of phosphorylation of the Ul-specific 70K protein is critical for its participation in a pre-catalytic step of the splicing reaction, which is vital for the successful splicing of pre-mRNA.
Journal of VirologyMolecular model of the A subunit of protein phosphatase 2A: interaction with other subunits and tumor antigens
139 Citations1994Ralf Ruediger, M Hentz +3 more
Native gel analysis of mutant A subunits synthesized in vitro demonstrated that the binding region for the B subunit, previously thought to include repeats 2 to 8, covers repeats 1 to 10 and that the B and C subunits cooperate in binding to the A subunit.
Proceedings of the National Academy of SciencesProtein composition of mammalian spliceosomes assembled in vitro.
114 Citations1990Robin Reed
An analysis of the protein composition of highly purified mammalian spliceosomes isolated by a two-step large-scale affinity chromatography procedure revealed a distinct set of pre-mRNA-specific proteins and a set of proteins that bind to pre- mRNA only in the presence of ATP.
Molecular and Cellular BiologyIdentification of Proteins That Interact with Exon Sequences, Splice Sites, and the Branchpoint Sequence during Each Stage of Spliceosome Assembly
111 Citations1996María‐Dolores Chiara, Or Gozani +4 more
The observation that these proteins are specifically associated with critical pre-mRNA sequence elements in functional complexes at different stages of spliceosome assembly implicates roles for these factors in key recognition events during the splicing pathway.
Nucleic Acids ResearchIdentification of an snRNP-associated kinase activity that phosphorylates arginine/serine rich domains typical of splicing factors
92 Citations1993Andreas Woppmann, Cindy L. Will +4 more
Tryptic phosphopeptide analysis revealed an overlapping set of at least four radiolabeled peptides in the in vivo and in vitro phosphorylated protein, suggesting that the snRNP-associated serine kinase is responsible, at least in part, for the 70K protein phosphorylation observed in vivo.
The EMBO JournalEvidence that U5 snRNP recognizes the 3′ splice site for catalytic step II in mammals
92 Citations1997María‐Dolores Chiara, Leon Palandjian +2 more
It is shown that U2AF65 crosslinking is replaced byCrosslinking of three proteins of 110, 116 and 220 kDa prior to catalytic step II, and evidence that all three proteins are components of U5 snRNP is provided, suggesting that U5SnRNP was positioned on the 3′ splice site by an interaction with U2 snR NP bound at the BPS and by a direct interaction with the pyrimidine tract.
ScienceCorrespondence Between a Mammalian Spliceosome Component and an Essential Yeast Splicing Factor
82 Citations1993Maria Bennett, Robin Reed
It is demonstrated that SAP 62 is the likely functional homolog of the yeast PRP11 protein, which associate stably with the spliceosome, contain a single zinc finger, and display significant amino acid sequence similarity.
Genes & DevelopmentCUS1, a suppressor of cold-sensitive U2 snRNA mutations, is a novel yeast splicing factor homologous to human SAP 145.
67 Citations1996Sandra E. Wells, Megan C. Neville +4 more
The yeast genome also encodes a homolog of human SAP 49, a protein that interacts strongly with both SAP 145 and pre-mRNA, underscoring the conservation of U2 snRNP proteins that function in spliceosome assembly.
PubMedThe [U4/U6.U5] tri-snRNP-specific 27K protein is a novel SR protein that can be phosphorylated by the snRNP-associated protein kinase.
60 Citations1997Sharon M. Fetzer, Jürgen Lauber +2 more
The tri-snRNP-specific 27K protein could potentially be involved in SR protein-mediated protein/protein interactions and, additionally, its phosphorylation state could modulate pre-mRNA splicing.