Identification of novel human WW domain-containing proteins by cloning of ligand targets.
A recently described protein module consisting of 35-40 semiconserved residues, termed the WW domain, has been identified in a number of diverse proteins including dystrophin and Yes-associated protein (YAP). Two putative ligands of YAP, termed WBP-1 and WBP-2, have been found previously to contain several short peptide regions consisting of PPPPY residues (PY motif) that mediate binding to the WW domain of YAP. Although the function(s) of the WW domain remain to be elucidated, these observations strongly support a role for the WW domain in protein-protein interactions. Here we report the isolation of three novel human cDNAs encoding a total of nine WW domains, using a newly developed approach termed COLT (cloning of ligand targets), in which the rapid cloning of modular protein domains is accomplished by screening cDNA expression libraries with specific peptide ligands. Two of the new genes identified appear to be members of a family of proteins, including Rsp5 and Nedd-4, which have ubiquitin-protein ligase activity. In addition, we demonstrate that peptides corresponding to PY and PY-like motifs present in several known signaling or regulatory proteins, including RasGAP, AP-2, p53BP-2 (p53-binding protein-2), interleukin-6 receptor-alpha, chloride channel CLCN5, and epithelial sodium channel ENaC, can selectively bind to certain of these novel WW domains.
Pirozzi
G
G
Cytogen Corporation, Princeton, New Jersey 08540-5309, USA. gpirozzi@mru3.cytogen.com
McConnell
S J
SJ
Uveges
A J
AJ
Carter
J M
JM
Sparks
A B
AB
Kay
B K
BK
Fowlkes
D M
DM
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D003001:Cloning, Molecular / Q000379:methods; D017124:Conserved Sequence; D018076:DNA, Complementary; D016189:Dystrophin / Q000737:chemistry*; D015723:Gene Library; D006801:Humans; D008024:Ligands; D008969:Molecular Sequence Data; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry
D018076:DNA, Complementary; D016189:Dystrophin; D008024:Ligands; D011993:Recombinant Fusion Proteins