Mutational analysis of human papillomavirus type 11 E5a oncoprotein.
In this study, we investigated the structural basis of human papillomavirus type 11 (HPV-11) E5a transforming activity at the amino acid level. The effects of insertion, deletion , and substitution mutations on teh E5a transforming activity were determined by the assay of anchorage-independent growth. In the conserved Cys-X-Cys structure, substitution of Ser for Cys-73 resulted in indistinguishable transforming activity, whereas substitution of Ser for Cys-75 or Ser for both Cys-73 and Cys-75 retained 50 and 42% transformation, respectively. This suggests that Cys at position 75 may be important for transformation. Charge and structural changes at teh COOH termini of several mutants impaired transformation significantly, but those at the middle region did so only mildly. In addition, the 16,000-molecular-weight pore-forming protein (16K protein) is known to associate with BPV-1, HPV-6, and HPV-16 E5 proteins. In this study, we investigated the correlation between E5a-16K binding affinity and the transforming activity of E5a by the use of 11 E5a mutants. Results show that E5a and these 11 E5a mutants could bind to the 16K protein when these proteins were coexpressed in COS cells, suggesting that simple binding of the 16K protein by E5a may not be sufficient for cell transformation.
Chen
S L
SL
Department of Microbiology and Immunology, National Defense Medical Center, Taipei, Taiwan, Republic of China.
Tsai
T Z
TZ
Han
C P
CP
Tsao
Y P
YP
D016475:3T3 Cells; D000251:Adenosine Triphosphatases / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002471:Cell Transformation, Neoplastic; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D009856:Oncogene Proteins, Viral / Q000737:chemistry* / Q000502:physiology; D027383:Papillomaviridae / Q000737:chemistry*; D013329:Structure-Activity Relationship; D014162:Transfection; D014617:Vacuoles / Q000201:enzymology
D009856:Oncogene Proteins, Viral; D000251:Adenosine Triphosphatases