SYNTHESIS, PHOSPHORYLATION, AND NUCLEAR-LOCALIZATION OF HUMAN PAPILLOMAVIRUS-E7 PROTEIN IN SCHIZOSACCHAROMYCES-POMBE

被引:29
作者
TOMMASINO, M
CONTORNI, M
SCARLATO, V
BUGNOLI, M
MAUNDRELL, K
CAVALIERI, F
机构
[1] SCLANO RES CTR,MOLEC VIROL,FIORENTINA 1,I-53100 SIENA,ITALY
[2] SCLAVO RES CTR,MOLEC BIOL,I-53100 SIENA,ITALY
关键词
antiserum; fission yeast; molecular cloning; Recombinant DNA; viral protein; yeast expression vector;
D O I
10.1016/0378-1119(90)90234-I
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
The complete E7 protein-encoding open reading frame of human papillomavirus type 16 (HPV-16) was expressed in the fission yeast Schizosaccharomyces pombe, under the control of a cloned yeast promoter. The HPV-16 E7 protein synthesized in S. pombe is a 17-kDa phosphoprotein which is recognized by anti-E7 antibodies (raised in rabbits against E7 fusion protein produced in Escherichia coli). The mobility during sodium dodecyl sulfate-polyacrylamide-gel electrophoresis of native E7 phosphoprotein synthesized in S. pombe is identical to that of the E7 phosphoprotein immunoprecipitated from human CaSki cells. Immunofluorescence staining showed that HPV-16 E7 phosphoprotein is localized in the nuclei of transformed S. pombe. These results indicate that E7 protein synthesized by S. pombe is apparently indistinguishable from HPV-16 protein synthesized in higher eukaryotic cells expressing genes of HPV-16, and also that the phosphorylated, nuclear HPV-16 E7 protein is synthesized in S. pombe in a form compatible with its biological activity. © 1990.
引用
收藏
页码:265 / 270
页数:6
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