Cooperatively between an upstream TATA-like sequence and a CAA repeated element mediates E1A-dependent negative repression of the H-2Kb class I gene.
In primary rodent cells transformed by the E1A region of the highly oncogenic adenovirus type 12, repression of transcription mediated by the far upstream TATA-like element was observed only in conjunction with either possible juxtaposition of a CAA repeated element in the presence of E1A and was dependent upon the relative arrangement of both the TATA-like and CAA repeated motifs in both homologous and heterologous promoter constructs. A gel shift competition study demonstrated that the TATA-binding protein (TBP) or a TBP-like protein can bind to both the upstream TATA-like sequence and the regular TATA box on the H-2Kb basal promoter. Moreover, employing immunoselection and cyclic amplification and selection of targets (CASTing) methods with nuclear extracts derived from Ad12-E1A transformants, we have identified a high affinity binding site in the H-2Kb class I promoter for E1A-associated DNA-binding proteins. The sequences of the binding sites were identified and were found to contain both the upstream TATA-like motif and the CAA repeated motifs. Our results suggest that the TATA-like sequence in the far upstream region of the H-2Kb gene is one of the elements that is required for Ad12-E1A-mediated negative repression.
Tsukuba Life Science Center, RIKEN (Institute of Physical and Chemical Research), Ibaraki, Japan.
X
X
Tang
H O
HO
Li
O
O
Sakatsume
T
T
Ohta
H
H
Tsutsui
A F
AF
Smit
M
M
Horikoshi
P
P
Kourilsky
A
A
Israël
G
G
Gachelin
D017500:Adenovirus E1A Proteins / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D001667:Binding, Competitive; D002472:Cell Transformation, Viral; D004268:DNA-Binding Proteins / Q000378:metabolism; D005786:Gene Expression Regulation; D006183:H-2 Antigens / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid; D012091:Repetitive Sequences, Nucleic Acid; D012097:Repressor Proteins / Q000502:physiology*; D016385:TATA Box; D014158:Transcription, Genetic
D017500:Adenovirus E1A Proteins; D004268:DNA-Binding Proteins; D006183:H-2 Antigens; C032876:H-2Kb protein, mouse; D012333:RNA, Messenger; D012097:Repressor Proteins