Cloning and characterization of the kinesin-related protein, Krp1p, in Schizosaccharomyces pombe.
Kinesin have been cloned in many organisms. They played important roles in the transport of cell organelles, polarized growth, and secretion. We report here the identification of a kinesin-related protein in Schizosaccharomyces pombe, which was named kinesin-related protein (Krplp). The primer sequences were driven from the highly conserved area of the kinesin genes in other organisms. We cloned kinesin genes from S. pombe using the PCR technique. Sequence analysis revealed that krp1+ has a 1,665 bp open-reading frame (ORF) that encoded a protein that consisted of 554 amino acids with a molecular weight of 61,900. It is homologous to the proteins that belong to the kinesin heavy chain (KHC) superfamily [GenBank accession No. AF156966 (genomic DNA) and AF247188 (mRNA)]. To characterize Krplp, the gene was disrupted and overexpressed in S. pombe. Cells that contained a krp1+ null allele were viable. Overexpression of Krp1p resulted in the inhibition of mitotic growth; cells became elongated, branched, and formed aberrant septa. To identify proteins that interact with Krplp, the yeast two-hybrid system was used. As a result, the novel protein, designated kinesin associated protein (Kap1p), was identified and showed structural homology to the proteins of the myosin family (GenBank accession No. AF351206). The data from the overexpression and two-hybrid study of Krplp may provide information that Krplp can have roles in cytokinesis with myosin.
Jeong
Jae Wook
JW
Division of Molecular Biology, Graduate School of Biotechnology, Korea University, Seoul.
Rhee
Dong Keun
DK
Cho
Soo Young
SY
Hae
Kwang Lae
KL
Kim
Dong Uk
DU
Won
Misun
M
Kim
Hyong Bai
HB
D001483:Base Sequence; D002455:Cell Division / Q000187:drug effects; D048429:Cell Size / Q000187:drug effects; D003001:Cloning, Molecular; D017353:Gene Deletion; D016547:Kinesins / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology; D008969:Molecular Sequence Data; D016296:Mutagenesis; D011485:Protein Binding; D012568:Schizosaccharomyces / Q000737:chemistry / Q000235:genetics*; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology; D017422:Sequence Analysis, DNA; D014161:Transduction, Genetic; D020798:Two-Hybrid System Techniques
D029702:Schizosaccharomyces pombe Proteins; C476402:Krp1 protein, S pombe; D016547:Kinesins