Functional analysis of human microtubule-based motor proteins, the kinesins and dyneins, in mitosis/cytokinesis using RNA interference.
Microtubule (MT)-based motor proteins, kinesins and dyneins, play important roles in multiple cellular processes including cell division. In this study, we describe the generation and use of an Escherichia coli RNase III-prepared human kinesin/dynein esiRNA library to systematically analyze the functions of all human kinesin/dynein MT motor proteins. Our results indicate that at least 12 kinesins are involved in mitosis and cytokinesis. Eg5 (a member of the kinesin-5 family), Kif2A (a member of the kinesin-13 family), and KifC1 (a member of the kinesin-14 family) are crucial for spindle formation; KifC1, MCAK (a member of the kinesin-13 family), CENP-E (a member of the kinesin-7 family), Kif14 (a member of the kinesin-3 family), Kif18 (a member of the kinesin-8 family), and Kid (a member of the kinesin-10 family) are required for chromosome congression and alignment; Kif4A and Kif4B (members of the kinesin-4 family) have roles in anaphase spindle dynamics; and Kif4A, Kif4B, MKLP1, and MKLP2 (members of the kinesin-6 family) are essential for cytokinesis. Using immunofluorescence analysis, time-lapse microscopy, and rescue experiments, we investigate the roles of these 12 kinesins in detail.
Zhu
Changjun
C
The Burnham Institute, La Jolla, CA 92037, USA.
Zhao
Jian
J
Bibikova
Marina
M
Leverson
Joel D
JD
Bossy-Wetzel
Ella
E
Fan
Jian-Bing
JB
Abraham
Robert T
RT
Jiang
Wei
W
D020413:3' Untranslated Regions; D000705:Anaphase; D001671:Biochemistry / Q000379:methods*; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry* / Q000378:metabolism; D002875:Chromosomes / Q000648:ultrastructure; D048749:Cytokinesis; D004398:Dyneins / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D015723:Gene Library; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D016547:Kinesins / Q000737:chemistry*; D018386:Kinetochores / Q000648:ultrastructure; D008856:Microscopy, Fluorescence; D008870:Microtubules / Q000737:chemistry* / Q000378:metabolism; D008938:Mitosis; D010641:Phenotype; D010957:Plasmids / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D008941:Spindle Apparatus; D013997:Time Factors; D014162:Transfection
D020413:3' Untranslated Regions; D002868:Chromosomal Proteins, Non-Histone; D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins; D004398:Dyneins; D016547:Kinesins