Functional analysis of the fission yeast Prp4 protein kinase involved in pre-mRNA splicing and isolation of a putative mammalian homologue

T Gross; M Lützelberger; H Weigmann; A Klingenhoff; S Shenoy; N F Käufer

doi:10.1093/nar/25.5.1028

Functional analysis of the fission yeast Prp4 protein kinase involved in pre-mRNA splicing and isolation of a putative mammalian homologue

Nucleic Acids Res. 1997 Mar 1;25(5):1028-35. doi: 10.1093/nar/25.5.1028.

Authors

T Gross¹, M Lützelberger, H Weigmann, A Klingenhoff, S Shenoy, N F Käufer

Affiliation

¹ Institüt für Genetik-Biozentrum, Technische Universität Braunschweig, Spielmannstrasse 7, D-38106 Braunschweig, Germany.

Abstract

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Animals
Binding Sites
DNA Primers
DNA, Complementary
Genetic Complementation Test
HeLa Cells
Humans
Mammals
Mice
Mitosis
Molecular Sequence Data
Mutagenesis, Site-Directed
Nuclear Proteins / metabolism
Protein Serine-Threonine Kinases / genetics
Protein Serine-Threonine Kinases / metabolism*
RNA Precursors / metabolism*
RNA Splicing Factors
RNA Splicing*
RNA-Binding Proteins
Rabbits
Recombinant Fusion Proteins / genetics
Recombinant Fusion Proteins / metabolism
Ribonucleoprotein, U4-U6 Small Nuclear / genetics
Ribonucleoprotein, U4-U6 Small Nuclear / metabolism
Schizosaccharomyces / enzymology*
Schizosaccharomyces / genetics
Schizosaccharomyces pombe Proteins*
Sequence Homology, Amino Acid
Serine-Arginine Splicing Factors

Substances

DNA Primers
DNA, Complementary
Nuclear Proteins
RNA Precursors
RNA Splicing Factors
RNA-Binding Proteins
Recombinant Fusion Proteins
Ribonucleoprotein, U4-U6 Small Nuclear
Schizosaccharomyces pombe Proteins
Serine-Arginine Splicing Factors
PRPF4B protein, human
Protein Serine-Threonine Kinases
Prpf4b protein, mouse
prp4 protein, S pombe