Accurate in vitro transcriptional initiation of the chick alpha 2 (Type I) collagen gene.
Chick genomic DNA containing the extreme 5' end of the alpha 2 (type I) collagen gene has been used as template in an in vitro HeLa cell transcription system. RNA polymerase II-dependent transcription initiates from a specific site on this DNA. The precise location of this site was determined by three types of experiments: sizing of in vitro-synthesized RNA runoff transcripts, comparing the sequence of the in vitro-made RNA transcripts with the structure of the DNA template, and identifying the first and second nucleotides of the in vitro-synthesized transcripts. Transcription was found to initiate 33 base pairs downstream from a canonical Goldberg-Hogness sequence (TATAAATA). This in vitro start site is the same as the initiation site of in vivo-synthesized collagen RNA.
D000818:Animals; D001483:Base Sequence; D002645:Chickens; D003001:Cloning, Molecular; D003094:Collagen / Q000235:genetics*; D004262:DNA Restriction Enzymes; D004274:DNA, Recombinant / Q000378:metabolism; D004278:DNA, Superhelical / Q000235:genetics; D005796:Genes; D006367:HeLa Cells / Q000378:metabolism; D010957:Plasmids; D006163:Ribonuclease T1; D013698:Templates, Genetic; D014158:Transcription, Genetic
D004274:DNA, Recombinant; D004278:DNA, Superhelical; D003094:Collagen; D004262:DNA Restriction Enzymes; D006163:Ribonuclease T1