Interactions of U24 from Roseolovirus with WW domains: canonical vs noncanonical.
U24 is a C-terminal membrane-anchored protein found in both human herpes virus type 6 and 7 (HHV-6 and HHV-7), with an N-terminal segment that is rich in prolines (PPxY motif in both HHV-6A and 7; PxxP motif in HHV-6A). Previous work has shown that U24 interacts strongly with Nedd4 WW domains, in particular, hNedd4L-WW3*. It was also shown that this interaction depends strongly on the nature of the amino acids that are upstream from the PY motif in U24. In this contribution, data was obtained from pull-downs, isothermal titration calorimetry, and NMR to further determine what modulates U24:WW domain interactions. Specifically, 3 non-canonical WW domains from human Smad ubiquitination regulatory factor (Smurf), namely hSmurf2-WW2, hSmurf2-WW3, and a tandem construct hSmurf2-WW2 + 3, were studied. Overall, the interactions between U24 and these Smurf WW domains were found to be weaker than those in U24:Nedd4 WW domain pairs, suggesting that U24 function is tightly linked to specific E3 ubiqitin ligases.
a Department of Chemistry, The University of British Columbia, Vancouver, BC V6T 1Z1, Canada.
Yurou
Y
Sang
a Department of Chemistry, The University of British Columbia, Vancouver, BC V6T 1Z1, Canada.
Rui
R
Zhang
b Michael Smith Laboratories and Department of Chemical and Biological Engineering, The University of British Columbia, Vancouver, BC V6T 1Z1, Canada.
A Louise
AL
Creagh
b Michael Smith Laboratories and Department of Chemical and Biological Engineering, The University of British Columbia, Vancouver, BC V6T 1Z1, Canada.
Charles A
CA
Haynes
a Department of Chemistry, The University of British Columbia, Vancouver, BC V6T 1Z1, Canada.
Suzana K
SK
Straus
D000595:Amino Acid Sequence; D056827:Endosomal Sorting Complexes Required for Transport / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D000075702:Nedd4 Ubiquitin Protein Ligases; D011485:Protein Binding; D011487:Protein Conformation; D000072417:Protein Domains; D019211:Roseolovirus / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D044767:Ubiquitin-Protein Ligases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D056827:Endosomal Sorting Complexes Required for Transport; D014764:Viral Proteins; D000075702:Nedd4 Ubiquitin Protein Ligases; C000618483:Nedd4 protein, human; C471641:SMURF2 protein, human; D044767:Ubiquitin-Protein Ligases
CTI; HHV-7); ITC; NMR; RMN; VHH-7); WW domains; domaines WW; human herpes virus type-6A and -7 (HHV-6A; interactions protéine–protéine; protein–protein interactions; virus de l’herpès humain de type 6A et -7 (VHH-6A