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PURIFICATION AND CHARACTERIZATION OF A KINASE SPECIFIC FOR THE SERINE-RICH AND ARGININE-RICH PRE-MESSENGER-RNA SPLICING FACTORS
JIAN-FANG GUI; HELENE TRONCHERE; SHARON D. CHANDLER; XIANG-DONG FU
1994
Source PublicationPROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume91Issue:23Pages:10824-10828
AbstractMembers of the SR family of pre-mRNA splicing factors are phosphoproteins that share a phosphoepitope specifically recognized by monoclonal antibody (mAb) 104. Recent studies have indicated that phosphorylation may regulate the activity and the intracellular localization of these splicing factors. Here, we report the purification and kinetic properties of SR protein kinase 1 (SRPK1), a kinase specific for SR family members. We demonstrate that the kinase specifically recognizes the SR domain, which contains serine/arginine repeats. Previous studies have shown that dephosphorylated SR proteins did not react with mAb 104 and migrated faster in SDS gels than SR proteins from mammalian cells. We show that SRPK1 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. Finally, we have correlated the effects of mutagenesis in the SR domain of SF2/ASF on splicing with those on phosphorylation of the protein by SRPK1, suggesting that phosphorylation of SR proteins is required for splicing.
KeywordPremessenger-rna Sr-proteins Conserved Family Site Selection Nuclear Phosphoproteins Factor Asf/sf2 Invitro Domains Enhancer Binding
DepartmentUNIV CALIF SAN DIEGO,DIV CELLULAR & MOLEC MED,LA JOLLA,CA 92093; UNIV CALIF SAN DIEGO,PROGRAM BIOMED SCI,LA JOLLA,CA 92093; CHINESE ACAD SCI,INST HYDROBIOL,WUHAN 430072,PEOPLES R CHINA
Subject AreaMultidisciplinary Sciences
Indexed BySCI
WOS IDWOS:A1994PQ93800015
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Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/152342/10312
Collection期刊论文
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JIAN-FANG GUI,HELENE TRONCHERE,SHARON D. CHANDLER,et al. PURIFICATION AND CHARACTERIZATION OF A KINASE SPECIFIC FOR THE SERINE-RICH AND ARGININE-RICH PRE-MESSENGER-RNA SPLICING FACTORS[J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA,1994,91(23):10824-10828.
APA JIAN-FANG GUI,HELENE TRONCHERE,SHARON D. CHANDLER,&XIANG-DONG FU.(1994).PURIFICATION AND CHARACTERIZATION OF A KINASE SPECIFIC FOR THE SERINE-RICH AND ARGININE-RICH PRE-MESSENGER-RNA SPLICING FACTORS.PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA,91(23),10824-10828.
MLA JIAN-FANG GUI,et al."PURIFICATION AND CHARACTERIZATION OF A KINASE SPECIFIC FOR THE SERINE-RICH AND ARGININE-RICH PRE-MESSENGER-RNA SPLICING FACTORS".PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 91.23(1994):10824-10828.
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