Altered motor activity of alternative splice variants of the mammalian kinesin-3 protein KIF1B.
Several mammalian kinesin motor proteins exist as multiple isoforms that arise from alternative splicing of a single gene. However, the roles of many motor protein splice variants remain unclear. The kinesin-3 motor protein KIF1B has alternatively spliced isoforms distinguished by the presence or absence of insertion sequences in the conserved amino-terminal region of the protein. The insertions are located in the loop region containing the lysine-rich cluster, also known as the K-loop, and in the hinge region adjacent to the motor domain. To clarify the functions of these alternative splice variants of KIF1B, we examined the biochemical properties of recombinant KIF1B with and without insertion sequences. In a microtubule-dependent ATPase assay, KIF1B variants that contained both insertions had higher activity and affinity for microtubules than KIF1B variants that contained no insertions. Mutational analysis of the K-loop insertion revealed that variants with a longer insertion sequence at this site had higher activity. However, the velocity of movement in motility assays was similar between KIF1B with and without insertion sequences. Our results indicate that splicing isoforms of KIF1B that vary in their insertion sequences have different motor activities.
Matsushita
Masafumi
M
Department of Biological Sciences, Graduate School of Science, Osaka University, Toyonaka, Osaka, Japan.
Yamamoto
Ruri
R
Mitsui
Keiji
K
Kanazawa
Hiroshi
H
D000251:Adenosine Triphosphatases / Q000378:metabolism; D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D054443:Cell Migration Assays / Q000379:methods*; D014644:Genetic Variation; D016547:Kinesins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008322:Mammals; D008870:Microtubules / Q000235:genetics / Q000378:metabolism; D020409:Molecular Motor Proteins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D020033:Protein Isoforms / Q000235:genetics; D017433:Protein Structure, Secondary / Q000235:genetics; D017434:Protein Structure, Tertiary / Q000235:genetics; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D018411:Spodoptera / Q000166:cytology / Q000378:metabolism
C490399:Kif1b protein, rat; D020409:Molecular Motor Proteins; D020033:Protein Isoforms; D011994:Recombinant Proteins; D000251:Adenosine Triphosphatases; D016547:Kinesins