Suppression of the bimC4 mitotic spindle defect by deletion of klpA, a gene encoding a KAR3-related kinesin-like protein in Aspergillus nidulans.
To investigate the relationship between structure and function of kinesin-like proteins, we have identified by polymerase chain reaction (PCR) a new kinesin-like protein in the filamentous fungus Aspergillus nidulans, which we have designated KLPA. DNA sequence analysis showed that the predicted KLPA protein contains a COOH terminal kinesin-like motor domain. Despite the structural similarity of KLPA to the KAR3 and NCD kinesin-like proteins of Saccharomyces cerevisiae and Drosophila melanogaster, which also posses COOH-terminal kinesin-like motor domains, there are no significant sequence similarities between the nonmotor or tail portions of these proteins. Nevertheless, expression studies in S. cerevisiae showed that klpA can complement a null mutation in KAR3, indicating that primary amino acid sequence conservation between the tail domains of kinesin-like proteins is not necessarily required for conserved function. Chromosomal deletion of the klpA gene exerted no observable mutant phenotype, suggesting that in A. nidulans there are likely to be other proteins functionally redundant with KLPA. Interestingly, the temperature sensitive phenotype of a mutation in another gene, bimC, which encodes a kinesin-like protein involved in mitotic spindle function in A. nidulans, was suppressed by deletion of klpA. We hypothesize that the loss of KLPA function redresses unbalanced forces within the spindle caused by mutation in bimC, and that the KLPA and BIMC kinesin-like proteins may play opposing roles in spindle function.
O'Connell
M J
MJ
Department of Pharmacology, UMDNJ-Robert Wood Johnson Medical School, Piscataway 08854-5635.
Meluh
P B
PB
Rose
M D
MD
Morris
N R
NR
D000595:Amino Acid Sequence; D001233:Aspergillus nidulans / Q000235:genetics* / Q000648:ultrastructure; D001483:Base Sequence; D003001:Cloning, Molecular; D017353:Gene Deletion; D015870:Gene Expression; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D016547:Kinesins / Q000235:genetics* / Q000502:physiology; D008938:Mitosis; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D016133:Polymerase Chain Reaction; D012331:RNA, Fungal / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D015183:Restriction Mapping; D008941:Spindle Apparatus / Q000648:ultrastructure
D009838:Oligodeoxyribonucleotides; D012331:RNA, Fungal; D012333:RNA, Messenger; D016547:Kinesins