Whole genome functional analysis identifies novel components required for mitotic spindle integrity in human cells.
BACKGROUND
The mitotic spindle is a complex mechanical apparatus required for accurate segregation of sister chromosomes during mitosis. We designed a genetic screen using automated microscopy to discover factors essential for mitotic progression. Using a RNA interference library of 49,164 double-stranded RNAs targeting 23,835 human genes, we performed a loss of function screen to look for small interfering RNAs that arrest cells in metaphase.


RESULTS
Here we report the identification of genes that, when suppressed, result in structural defects in the mitotic spindle leading to bent, twisted, monopolar, or multipolar spindles, and cause cell cycle arrest. We further describe a novel analysis methodology for large-scale RNA interference datasets that relies on supervised clustering of these genes based on Gene Ontology, protein families, tissue expression, and protein-protein interactions.


CONCLUSIONS
This approach was utilized to classify functionally the identified genes in discrete mitotic processes. We confirmed the identity for a subset of these genes and examined more closely their mechanical role in spindle architecture.
Rines
Daniel R
DR
Genomics Institute of Novartis Research Foundation, John Jay Hopkins Drive, San Diego, California 92121, USA. drines@gnf.org
Gomez-Ferreria
Maria Ana
MA
Zhou
Yingyao
Y
DeJesus
Paul
P
Grob
Seanna
S
Batalov
Serge
S
Labow
Marc
M
Huesken
Dieter
D
Mickanin
Craig
C
Hall
Jonathan
J
Reinhardt
Mischa
M
Natt
Francois
F
Lange
Joerg
J
Sharp
David J
DJ
Chanda
Sumit K
SK
Caldwell
Jeremy S
JS
D015894:Genome, Human; D006801:Humans; D008938:Mitosis / Q000235:genetics*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics* / Q000502:physiology*; D008941:Spindle Apparatus / Q000378:metabolism* / Q000648:ultrastructure*
D034741:RNA, Small Interfering