RNA interference-mediated silencing of mitotic kinesin KIF14 disrupts cell cycle progression and induces cytokinesis failure.
KIF14 is a microtubule motor protein whose elevated expression is associated with poor-prognosis breast cancer. Here we demonstrate KIF14 accumulation in mitotic cells, where it associated with developing spindle poles and spindle microtubules. Cells at later stages of mitosis were characterized by the concentration of KIF14 at the midbody. Time-lapse microscopy revealed that strong RNA interference (RNAi)-mediated silencing of KIF14 induced cytokinesis failure, causing several rounds of endoreduplication and resulting in multinucleated cells. Additionally, less efficacious KIF14-specific short interfering RNAs (siRNAs) induced multiple phenotypes, all of which resulted in acute apoptosis. Our data demonstrate the ability of siRNA-mediated silencing to generate epiallelic hypomorphs associated with KIF14 depletion. Furthermore, the link we observed between siRNA efficacy and phenotypic outcome indicates that distinct stages during cell cycle progression are disrupted by the differential modulation of KIF14 expression.
Carleton
Michael
M
Rosetta Inpharmatics LLC, 401 Terry Ave., N. Seattle, Washington 98109, USA. michael_carleton@merck.com
Mao
Mao
M
Biery
Matthew
M
Warrener
Paul
P
Kim
Sammy
S
Buser
Carolyn
C
Marshall
C Gary
CG
Fernandes
Christine
C
Annis
James
J
Linsley
Peter S
PS
D000251:Adenosine Triphosphatases / Q000032:analysis; D000595:Amino Acid Sequence; D017209:Apoptosis; D002453:Cell Cycle / Q000502:physiology*; D002460:Cell Line; D003001:Cloning, Molecular; D016384:Consensus Sequence; D048749:Cytokinesis / Q000502:physiology*; D004815:Epidermal Growth Factor / Q000378:metabolism; D005456:Fluorescent Dyes; D015972:Gene Expression Regulation, Neoplastic; D020868:Gene Silencing; D045325:HCT116 Cells; D006367:HeLa Cells; D006801:Humans; D015151:Immunoblotting; D007211:Indoles; D016547:Kinesins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D018715:Microscopy, Video; D008969:Molecular Sequence Data; D015513:Oncogene Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019683:Penetrance; D017434:Protein Structure, Tertiary; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics
D005456:Fluorescent Dyes; D007211:Indoles; D015513:Oncogene Proteins; D034741:RNA, Small Interfering; C007293:DAPI; D004815:Epidermal Growth Factor; D000251:Adenosine Triphosphatases; C502598:KIF14 protein, human; D016547:Kinesins