Expression of the potential novel gene E6DG1 downregulated by the E6 protein of human papillomavirus type 16 is correlated with anchorage-independent growth.
A novel gene transcript that is downregulated by HPV16 E6 protein was identified in mouse cells using differential hybridization and designated E6DG1. The cloned cDNA of E6DG1 was 1.3 kb in length and contained a small ORF potentially encoding a polypeptide of 45 amino acids. In vitro transcription and translation of E6DG1 cDNA resulted in a product of approximately 7 kDa and Western blot analysis using antibodies for E6DG1 peptide detected 7 and 14 kDa proteins. Downregulation of E6DG1 mRNA levels in cells expressing HPV16 E6 protein was observed at subconfluent cell densities, but not in confluent cells. Repression of E6DG1 protein enhanced the anchorage-independent growth and weakened the cell adhesion in Panc1 cells. Immunofluorescence analysis revealed the localization of E6DG1 within the nucleus. These results indicate that the E6DG1 protein may function in a signaling pathway related to anchorage-independent growth and adhesion control.
Sakao
Seiichiro
S
Department of Molecular Virology, Chiba University School of Medicine, Chiba 260-8670, Japan.
Hashimoto
Tomohiro
T
Shino
Yuji
Y
Saito
Takeshi
T
Kado
Satoshi
S
Sashiyama
Hiroshi
H
Tatsumi
Koichiro
K
Kuriyama
Takayuki
T
Shirasawa
Hiroshi
H
D000256:Adenoviridae; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D002448:Cell Adhesion / Q000502:physiology*; D002452:Cell Count; D002455:Cell Division / Q000502:physiology*; D002460:Cell Line; D003001:Cloning, Molecular; D015536:Down-Regulation; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009856:Oncogene Proteins, Viral / Q000502:physiology*; D027383:Papillomaviridae / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism; D011817:Rabbits; D012097:Repressor Proteins; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
C068651:E6 protein, Human papillomavirus type 16; C470511:E6DG1 protein, mouse; D009856:Oncogene Proteins, Viral; D012333:RNA, Messenger; D012097:Repressor Proteins; D014157:Transcription Factors; D016159:Tumor Suppressor Protein p53