The role of the coiled-coil motif in interactions mediated by TPD52.
TPD52 (D52)-like proteins are small coiled-coil motif-bearing proteins first identified through their expression in human breast carcinoma that mutually interact in hetero- and homomeric fashions. However, it has been unclear whether the coiled-coil motif is sufficient, or even necessary, for these interactions to occur. We have therefore examined the binding activities of a panel of C-terminally deleted D52 proteins in both the yeast two-hybrid system and pull-down assays. In the yeast two-hybrid system, interactions were only detected when regions C-terminal to the coiled-coil motif were also present. However, using pull-down assays, interactions were detected for all deletion mutants which included the coiled-coil motif. This suggests that the coiled-coil motif is indeed necessary for interactions mediated by D52 proteins, but that C-terminal protein regions facilitate and/or stabilize these interactions.
Sathasivam
P
P
Molecular Oncology Laboratory, The Children's Hospital at Westmead, Westmead 2145, New South Wales, Australia.
Bailey
A M
AM
Crossley
M
M
Byrne
J A
JA
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D014644:Genetic Variation; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D020798:Two-Hybrid System Techniques
D009363:Neoplasm Proteins; D011993:Recombinant Fusion Proteins; C114751:TPD52 protein, human; C486415:Tpd52 protein, mouse