Binding of pleckstrin homology domains to WD40/beta-transducin repeat containing segments of the protein product of the Lis-1 gene.
In previous experiments we demonstrated an interaction between certain pleckstrin homology (PH) domains and regions containing the so-called WD40 (or beta-transducin) repeats of the beta subunit of trimeric G-proteins (G beta), a finding we here extend to the PH domains of the src-related tyrosine kinase TecIIa and the GTPase dynamin. To examine the possibility that WD40 repeats in molecules other than G beta might also bind PH domains we examined PAFAH-45, the protein product of the Lis-1 gene, which contains 7 WD40 repeats. We found that 1) Purified PAFAH-45 binds PH domain constructs in vitro. 2) Protein constructs expressing all 7 WD40 repeats of PAFAH-45 but lacking the N-terminal non WD40 region also bind PH domains of beta-adrenergic receptor kinase, beta-spectrin, TecIIa and dynamin but with a differing hierarchy of affinities than that seen with G beta. 3). PAFAH-45 WD40 repeats will reduce the binding of PH domains to brain G beta and brain G beta gamma will reduce the binding of PH domains to PAFAH-45. These data support the hypothesis that PH domain/WD40 interactions are involved in a wide variety of important protein/protein interactions.
Wang
D S
DS
University of Florida College of Medicine, Department of Neuroscience, Gainesville, Florida 32610, USA.
Shaw
R
R
Hattori
M
M
Arai
H
H
Inoue
K
K
Shaw
G
G
D000818:Animals; D001483:Base Sequence; D001667:Binding, Competitive; D001798:Blood Proteins / Q000737:chemistry; D002417:Cattle; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D034281:Dynamins; D020558:GTP Phosphohydrolases / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000378:metabolism; D066298:In Vitro Techniques; D046911:Macromolecular Substances; D008869:Microtubule-Associated Proteins; D008969:Molecular Sequence Data; D010750:Phosphoproteins; D011485:Protein Binding; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D012091:Repetitive Sequences, Nucleic Acid; D015398:Signal Transduction; D013049:Spectrin / Q000378:metabolism; D015293:Transducin / Q000737:chemistry; D051552:beta-Adrenergic Receptor Kinases
D001798:Blood Proteins; D046911:Macromolecular Substances; D008869:Microtubule-Associated Proteins; D010750:Phosphoproteins; D011506:Proteins; C039077:platelet protein P47; D013049:Spectrin; D011505:Protein-Tyrosine Kinases; D017868:Cyclic AMP-Dependent Protein Kinases; D051552:beta-Adrenergic Receptor Kinases; D020558:GTP Phosphohydrolases; D019204:GTP-Binding Proteins; D015293:Transducin; D034281:Dynamins