TY - JOUR
T1 - Functional analysis of the nucleotide sequence surrounding the cap site for adenovirus type 5 region E1A messenger RNAs
AU - Hearing, Patrick
AU - Shenk, Thomas
AU - Chambon, P.
N1 - Funding Information:
We acknowledge the competent technical assistance of Martha Marlow. This work was supported by a grant (MV-45) from the American Cancer Society. P.H. is a Fellow of the Jane Coffin Childs Memorial Fund for Medical Research; T.S. was an Established Investigator of the American Heart Association.
PY - 1983/7/15
Y1 - 1983/7/15
N2 - We have constructed a set of small, dispersed deletion mutations in the sequences surrounding the cap site of the adenovirus type 5 region E1A transcription unit. The effects of the mutations on E1A transcription were studied in vitro using a HeLa cell-free extract, and in vivo by reconstructing the mutations back into intact viral chromosomes and analyzing E1A messenger RNAs synthesized after infection of HeLa cells. The sequence between -35 and +20 (relative to the cap site at +1) was important for efficient E1A transcription and cap site selection in vitro. This region includes the "TATA homology, which appeared essential for transcription. Sequences upstream of -35 were dispensable for transcription in vitro. Different results were found upon analysis of the same set of deletions in vivo. None of the mutations affected the steady-state levels of cytoplasmic, E1A-specific mRNAs found in infected HeLa cells by more than twofold. Deletions of the TATA homology, however, generated E1A mRNAs with heterogeneous 5′ ends, and deletions downstream of the homology displaced the 5′ end of mRNAs by about the size of the deletion.
AB - We have constructed a set of small, dispersed deletion mutations in the sequences surrounding the cap site of the adenovirus type 5 region E1A transcription unit. The effects of the mutations on E1A transcription were studied in vitro using a HeLa cell-free extract, and in vivo by reconstructing the mutations back into intact viral chromosomes and analyzing E1A messenger RNAs synthesized after infection of HeLa cells. The sequence between -35 and +20 (relative to the cap site at +1) was important for efficient E1A transcription and cap site selection in vitro. This region includes the "TATA homology, which appeared essential for transcription. Sequences upstream of -35 were dispensable for transcription in vitro. Different results were found upon analysis of the same set of deletions in vivo. None of the mutations affected the steady-state levels of cytoplasmic, E1A-specific mRNAs found in infected HeLa cells by more than twofold. Deletions of the TATA homology, however, generated E1A mRNAs with heterogeneous 5′ ends, and deletions downstream of the homology displaced the 5′ end of mRNAs by about the size of the deletion.
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U2 - 10.1016/S0022-2836(83)80112-0
DO - 10.1016/S0022-2836(83)80112-0
M3 - Article
C2 - 6876165
AN - SCOPUS:0021104422
SN - 0022-2836
VL - 167
SP - 809
EP - 822
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
IS - 4
ER -