'Edgetic' perturbation of a C. elegans BCL2 ortholog.
Genes and gene products do not function in isolation but within highly interconnected 'interactome' networks, modeled as graphs of nodes and edges representing macromolecules and interactions between them, respectively. We propose to investigate genotype-phenotype associations by methodical use of alleles that lack single interactions, while retaining all others, in contrast to genetic approaches designed to eliminate gene products completely. We describe an integrated strategy based on the reverse yeast two-hybrid system to isolate and characterize such edge-specific, or 'edgetic', alleles. We established a proof of concept with CED-9, a Caenorhabditis elegans BCL2 ortholog. Using ced-9 edgetic alleles, we uncovered a new potential functional link between apoptosis and a centrosomal protein. This approach is amenable to higher throughput and is particularly applicable to interactome network analysis in organisms for which transgenesis is straightforward.
Center for Cancer Systems Biology (CCSB) and Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, Massachusetts, USA.
Matija
M
Dreze
Benoit
B
Charloteaux
Stuart
S
Milstein
Pierre-Olivier
PO
Vidalain
Muhammed A
MA
Yildirim
Quan
Q
Zhong
Nenad
N
Svrzikapa
Viviana
V
Romero
Géraldine
G
Laloux
Robert
R
Brasseur
Jean
J
Vandenhaute
Mike
M
Boxem
Michael E
ME
Cusick
David E
DE
Hill
Marc
M
Vidal
D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D017173:Caenorhabditis elegans / Q000235:genetics; D029742:Caenorhabditis elegans Proteins / Q000235:genetics* / Q000502:physiology; D002135:Calcium-Binding Proteins / Q000235:genetics; D017238:Genes, Helminth; D005838:Genotype; D008958:Models, Molecular; D010641:Phenotype; D025941:Protein Interaction Mapping / Q000379:methods*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics*; D012097:Repressor Proteins / Q000502:physiology; D020798:Two-Hybrid System Techniques
D029742:Caenorhabditis elegans Proteins; D002135:Calcium-Binding Proteins; C089224:Ced-4 protein, C elegans; C087233:Ced-9 protein, C elegans; C112486:EGL-1 protein, C elegans; D019253:Proto-Oncogene Proteins c-bcl-2; D012097:Repressor Proteins