PMID- 10196197
OWN - NLM
STAT- MEDLINE
DCOM- 19990517
LR  - 20190508
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 16
DP  - 1999 Apr 16
TI  - The subcellular localization of SF2/ASF is regulated by direct interaction with
      SR protein kinases (SRPKs).
PG  - 11125-31
AB  - Serine/arginine-rich (SR) proteins play an important role in constitutive and
      alternative pre-mRNA splicing. The C-terminal arginine-serine domain of these
      proteins, such as SF2/ASF, mediates protein-protein interactions and is
      phosphorylated in vivo. Using glutathione S-transferase (GST)-SF2/ASF-affinity
      chromatography, the SF2/ASF kinase activity was co-purified from HeLa cells with 
      a 95-kDa protein, which was recognized by an anti-SR protein kinase (SRPK) 1
      monoclonal antibody. Recombinant SRPK1 and SRPK2 bound to and phosphorylated
      GST-SF2/ASF in vitro. Phosphopeptide mapping showed that identical sites were
      phosphorylated in the pull-down kinase reaction with HeLa extracts and by
      recombinant SRPKs. Epitope-tagged SF2/ASF transiently expressed in COS7 cells
      co-immunoprecipitated with SRPKs. Deletion analysis mapped the phosphorylation
      sites to a region containing an (Arg-Ser)8 repeat beginning at residue 204, and
      far-Western analysis showed that the region is required for binding of SRPKs to
      SF2/ASF. Further binding studies showed that SRPKs bound unphosphorylated SF2/ASF
      but did not bind phosphorylated SF2/ASF. Expression of an SRPK2 kinase-inactive
      mutant caused accumulation of SF2/ASF in the cytoplasm. These results suggest
      that the formation of complexes between SF2/ASF and SRPKs, which is influenced by
      the phosphorylation state of SF2/ASF, may have regulatory roles in the assembly
      and localization of this splicing factor.
FAU - Koizumi, J
AU  - Koizumi J
AD  - Department of Functional Genomics, Medical Research Institute, Tokyo Medical and 
      Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113, Japan.
FAU - Okamoto, Y
AU  - Okamoto Y
FAU - Onogi, H
AU  - Onogi H
FAU - Mayeda, A
AU  - Mayeda A
FAU - Krainer, A R
AU  - Krainer AR
FAU - Hagiwara, M
AU  - Hagiwara M
LA  - eng
GR  - CA13106/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Nuclear Proteins)
RN  - 0 (RNA-Binding Proteins)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 170974-22-8 (Serine-Arginine Splicing Factors)
RN  - EC 2.5.1.18 (Glutathione Transferase)
RN  - EC 2.7.1.- (SRPK1 protein, human)
RN  - EC 2.7.11.1 (Protein-Serine-Threonine Kinases)
SB  - IM
MH  - Glutathione Transferase/metabolism
MH  - HeLa Cells
MH  - Humans
MH  - Nuclear Proteins/*metabolism
MH  - Phosphorylation
MH  - Protein-Serine-Threonine Kinases/*metabolism
MH  - RNA Splicing
MH  - RNA-Binding Proteins
MH  - Recombinant Fusion Proteins/metabolism
MH  - Serine-Arginine Splicing Factors
MH  - Subcellular Fractions/*metabolism
EDAT- 1999/04/10 00:00
MHDA- 1999/04/10 00:01
CRDT- 1999/04/10 00:00
PHST- 1999/04/10 00:00 [pubmed]
PHST- 1999/04/10 00:01 [medline]
PHST- 1999/04/10 00:00 [entrez]
AID - 10.1074/jbc.274.16.11125 [doi]
PST - ppublish
SO  - J Biol Chem. 1999 Apr 16;274(16):11125-31. doi: 10.1074/jbc.274.16.11125.