Functional analysis of the fission yeast Prp4 protein kinase involved in pre-mRNA splicing and isolation of a putative mammalian homologue.
The prp4 gene of Schizosaccharomyces pombe encodes a protein kinase. A physiological substrate is not yet known. A mutational analysis of prp4 revealed that the protein consists of a short N-terminal domain, containing several essential motifs, which is followed by the kinase catalytic domain comprising the C-terminus of the protein. Overexpression of N-terminal mutations disturbs mitosis and produces elongated cells, Using a PCR approach, we isolated a putative homologue of Prp4 from human and mouse cells. The mammalian kinase domain is 53% identical to the kinase domain of Prp4. The short N-terminal domains share <20% identical amino acids, but contain conserved motifs. A fusion protein consisting of the N-terminal region from S. pombe followed by the mammalian kinase domain complements a temperature-sensitive prp4 mutation of S. pombe. Prp4 and the recombinant yeast/mouse protein kinase phosphorylate the human SR splicing factor ASF/SF2 in vitro in its RS domain.
Gross
T
T
Institüt für Genetik-Biozentrum, Technische Universität Braunschweig, Spielmannstrasse 7, D-38106 Braunschweig, Germany.
Lützelberger
M
M
Weigmann
H
H
Klingenhoff
A
A
Shenoy
S
S
Käufer
N F
NF
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D017931:DNA Primers; D018076:DNA, Complementary; D005816:Genetic Complementation Test; D006367:HeLa Cells; D006801:Humans; D008322:Mammals; D051379:Mice; D008938:Mitosis; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009687:Nuclear Proteins / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D012322:RNA Precursors / Q000378:metabolism*; D012326:RNA Splicing; D000072260:RNA Splicing Factors; D016601:RNA-Binding Proteins; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017414:Ribonucleoprotein, U4-U6 Small Nuclear / Q000235:genetics / Q000378:metabolism; D012568:Schizosaccharomyces / Q000201:enzymology* / Q000235:genetics; D029702:Schizosaccharomyces pombe Proteins; D017386:Sequence Homology, Amino Acid; D000068103:Serine-Arginine Splicing Factors
D017931:DNA Primers; D018076:DNA, Complementary; D009687:Nuclear Proteins; D012322:RNA Precursors; D000072260:RNA Splicing Factors; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D017414:Ribonucleoprotein, U4-U6 Small Nuclear; D029702:Schizosaccharomyces pombe Proteins; D000068103:Serine-Arginine Splicing Factors; C489984:PRPF4B protein, human; D017346:Protein Serine-Threonine Kinases; C489983:Prpf4b protein, mouse; C489981:prp4 protein, S pombe