Cloning and characterization of an alternatively spliced form of SR protein kinase 1 that interacts specifically with scaffold attachment factor-B.
Serine/arginine protein kinases have been conserved throughout evolution and are thought to play important roles in the regulation of mRNA processing, nuclear import, germline development, polyamine transport, and ion homeostasis. Human SRPK1, which was first identified as a kinase specific for the SR family of splicing factors, is located on chromosome 6p21.2-p21.3. We report here the cloning and characterization of SRPK1a, which is encoded by an alternatively processed transcript derived from the SRPK1 gene. SRPK1a contains an insertion of 171 amino acids at its NH(2)-terminal domain and is similar to SRPK1 in substrate specificity and subcellular localization. Moreover, both isoforms can induce alternative splicing of human tau exon 10 in transfected cells. Using the yeast two-hybrid assay, we found that the extended NH(2)-terminal domain of SRPK1a interacts with Scaffold Attachment Factor-B, a nuclear scaffold-associated protein. Confirmation of this interaction was provided by in vitro binding assays, as well as by co-immunoprecipitation from 293T cells doubly transfected with SRPK1a and SAF-B. Our studies suggest that different SRPK family members are uniquely regulated and targeted and thus the multiple SRPK kinases present in higher eukaryotes may perform specialized and differentiable functions.
Nikolakaki
E
E
Laboratory of Biochemistry, School of Chemistry, The Aristotelian University of Thessaloniki, Thessaloniki 54006, Greece. nikol@ccf.auth.gr
Kohen
R
R
Hartmann
A M
AM
Stamm
S
S
Georgatsou
E
E
Giannakouros
T
T
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002451:Cell Compartmentation; D003001:Cloning, Molecular; D019143:Evolution, Molecular; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D011485:Protein Binding; D020033:Protein Isoforms; D017346:Protein Serine-Threonine Kinases / Q000235:genetics* / Q000378:metabolism*; D021382:Protein Sorting Signals; D051381:Rats; D012261:Ribonucleoproteins / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D013737:Testis / Q000737:chemistry; D014018:Tissue Distribution; D020798:Two-Hybrid System Techniques
D034441:Heterogeneous-Nuclear Ribonucleoproteins; D020033:Protein Isoforms; D021382:Protein Sorting Signals; D012261:Ribonucleoproteins; C087728:SRPK1 protein, human; D017346:Protein Serine-Threonine Kinases