The E7 oncoprotein of human papillomavirus type 16 interacts with F-actin in vitro and in vivo.
We report here that E7 oncoprotein of human papillomavirus type 16 (HPV-16) forms a complex in vivo and in vitro with actin, one of the components of the cellular cytoskeleton. The in vivo interaction was detected by immunofluorescent staining and confocal microscopic examination of normal human oral keratinocytes (NHOK) and CV-1 cells after transient expression of E7 employing the vaccinia virus-T7 RNA polymerase system and by coimmunoprecipitation from an immortalized, nontumorigenic cell line obtained after transfecting NHOK with the cloned HPV-16 DNA genome. The in vitro interaction was detected by cosedimentation of bacterially expressed E7 phosphorylated with rabbit reticulocyte lysate or purified casein kinase II (CKII) prior to incubation with F-actin. This interaction was inhibited if E7 phosphorylation by the rabbit reticulocyte lysate was prevented with heparin, a CKII inhibitor, or if the amino acids Ser-31 and Ser-32 in E7, which are phosphorylated by CKII, were replaced with amino acids that cannot be phosphorylated. Interestingly, a decrease in the amount of polymerized actin occurred in cells expressing E7.
Rey
O
O
School of Dentistry, University of California, Los Angeles, Los Angeles, California, 90095-1668, USA.
Lee
S
S
Baluda
M A
MA
Swee
J
J
Ackerson
B
B
Chiu
R
R
Park
N H
NH
D000199:Actins / Q000378:metabolism*; D000818:Animals; D001704:Biopolymers / Q000378:metabolism; D002460:Cell Line; D002461:Cell Line, Transformed; D006801:Humans; D007424:Intracellular Fluid / Q000378:metabolism; D015603:Keratinocytes / Q000201:enzymology / Q000821:virology; D009856:Oncogene Proteins, Viral / Q000096:biosynthesis / Q000378:metabolism*; D027383:Papillomaviridae / Q000201:enzymology / Q000378:metabolism*; D050725:Papillomavirus E7 Proteins; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000096:biosynthesis / Q000378:metabolism; D011817:Rabbits
D000199:Actins; D001704:Biopolymers; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; C059731:oncogene protein E7, Human papillomavirus type 16; D011505:Protein-Tyrosine Kinases