Systematic interactome mapping and genetic perturbation analysis of a C. elegans TGF-beta signaling network.
To initiate a system-level analysis of C. elegans DAF-7/TGF-beta signaling, we combined interactome mapping with single and double genetic perturbations. Yeast two-hybrid (Y2H) screens starting with known DAF-7/TGF-beta pathway components defined a network of 71 interactions among 59 proteins. Coaffinity purification (co-AP) assays in mammalian cells confirmed the overall quality of this network. Systematic perturbations of the network using RNAi, both in wild-type and daf-7/TGF-beta pathway mutant animals, identified nine DAF-7/TGF-beta signaling modifiers, seven of which are conserved in humans. We show that one of these has functional homology to human SNO/SKI oncoproteins and that mutations at the corresponding genetic locus daf-5 confer defects in DAF-7/TGF-beta signaling. Our results reveal substantial molecular complexity in DAF-7/TGF-beta signal transduction. Integrating interactome maps with systematic genetic perturbations may be useful for developing a systems biology approach to this and other signaling modules.
Tewari
Muneesh
M
Center for Cancer Systems Biology, Dana-Farber Cancer Institute, Boston, Massachusetts 02115, USA.
Hu
Patrick J
PJ
Ahn
Jin Sook
JS
Ayivi-Guedehoussou
Nono
N
Vidalain
Pierre-Olivier
PO
Li
Siming
S
Milstein
Stuart
S
Armstrong
Chris M
CM
Boxem
Mike
M
Butler
Maurice D
MD
Busiguina
Svetlana
S
Rual
Jean-François
JF
Ibarrola
Nieves
N
Chaklos
Sabrina T
ST
Bertin
Nicolas
N
Vaglio
Philippe
P
Edgley
Mark L
ML
King
Kevin V
KV
Albert
Patrice S
PS
Vandenhaute
Jean
J
Pandey
Akhilesh
A
Riddle
Donald L
DL
Ruvkun
Gary
G
Vidal
Marc
M
D000818:Animals; D017173:Caenorhabditis elegans / Q000235:genetics / Q000378:metabolism*; D029742:Caenorhabditis elegans Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D002874:Chromosome Mapping; D003360:Cosmids; D018507:Gene Expression Regulation, Developmental; D017238:Genes, Helminth; D023281:Genomics; D006801:Humans; D007668:Kidney; D008954:Models, Biological; D009154:Mutation; D016366:Open Reading Frames; D020543:Proteome / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000378:metabolism; D015398:Signal Transduction; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000235:genetics / Q000378:metabolism*; D019076:Transgenes; D020798:Two-Hybrid System Techniques
D029742:Caenorhabditis elegans Proteins; D020543:Proteome; D034741:RNA, Small Interfering; D016212:Transforming Growth Factor beta