Abstract
Transcription of the carbamoyl-phosphate synthase (glutamine- hydrolyzing)/aspartate carbamoyltransferase/dihydroorotase (CAD) gene from the Syrian hamster, Mesocricetus auratus, starts at a single major site. We characterized the cis-acting elements that position RNA polymerase II at the correct start site in the CAD promoter. Sequence alignment showed that the CAD promoter lacks a TATA box, but contains a consensus initiator. Mutational analysis of the CAD promoter demonstrated that the sequences between -81 and +26 were sufficient for accurate and efficient transcription in vitro and in vivo; binding sites for the transcription factor Sp1 around -70 and -49 were necessary for transcriptional activity. The binding site at -49 directed initiation about 50 base pairs downstream. A ubiquitous activator protein, Honk, bound to the CAD promoter between -30 and -12, but did not participate in start site selection. The sequences around +1, which contain the consensus initiator, contributed to promoter activity; however, the presence of a consensus initiator in this region was neither necessary nor sufficient for transcription. We concluded from these results that the Sp1 binding site at - 49 substituted for the missing TATA box and played a major role in start site selection at the CAD promoter.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 2252-2257 |
| Number of pages | 6 |
| Journal | Journal of Biological Chemistry |
| Volume | 269 |
| Issue number | 3 |
| State | Published - Jan 21 1994 |
| Externally published | Yes |
Fingerprint
ASJC Scopus subject areas
- Biochemistry
Cite this
Start site selection at the TATA-less carbamoyl-phosphate synthase (glutamine-hydrolyzing)/aspartate carbamoyltransferase/dihydroorotase promoter. / Kollmar, Richard; Sukow, Kristine A.; Sponagle, Stephen K.; Farnham, Peggy J.
In: Journal of Biological Chemistry, Vol. 269, No. 3, 21.01.1994, p. 2252-2257.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Start site selection at the TATA-less carbamoyl-phosphate synthase (glutamine-hydrolyzing)/aspartate carbamoyltransferase/dihydroorotase promoter
AU - Kollmar, Richard
AU - Sukow, Kristine A.
AU - Sponagle, Stephen K.
AU - Farnham, Peggy J.
PY - 1994/1/21
Y1 - 1994/1/21
N2 - Transcription of the carbamoyl-phosphate synthase (glutamine- hydrolyzing)/aspartate carbamoyltransferase/dihydroorotase (CAD) gene from the Syrian hamster, Mesocricetus auratus, starts at a single major site. We characterized the cis-acting elements that position RNA polymerase II at the correct start site in the CAD promoter. Sequence alignment showed that the CAD promoter lacks a TATA box, but contains a consensus initiator. Mutational analysis of the CAD promoter demonstrated that the sequences between -81 and +26 were sufficient for accurate and efficient transcription in vitro and in vivo; binding sites for the transcription factor Sp1 around -70 and -49 were necessary for transcriptional activity. The binding site at -49 directed initiation about 50 base pairs downstream. A ubiquitous activator protein, Honk, bound to the CAD promoter between -30 and -12, but did not participate in start site selection. The sequences around +1, which contain the consensus initiator, contributed to promoter activity; however, the presence of a consensus initiator in this region was neither necessary nor sufficient for transcription. We concluded from these results that the Sp1 binding site at - 49 substituted for the missing TATA box and played a major role in start site selection at the CAD promoter.
AB - Transcription of the carbamoyl-phosphate synthase (glutamine- hydrolyzing)/aspartate carbamoyltransferase/dihydroorotase (CAD) gene from the Syrian hamster, Mesocricetus auratus, starts at a single major site. We characterized the cis-acting elements that position RNA polymerase II at the correct start site in the CAD promoter. Sequence alignment showed that the CAD promoter lacks a TATA box, but contains a consensus initiator. Mutational analysis of the CAD promoter demonstrated that the sequences between -81 and +26 were sufficient for accurate and efficient transcription in vitro and in vivo; binding sites for the transcription factor Sp1 around -70 and -49 were necessary for transcriptional activity. The binding site at -49 directed initiation about 50 base pairs downstream. A ubiquitous activator protein, Honk, bound to the CAD promoter between -30 and -12, but did not participate in start site selection. The sequences around +1, which contain the consensus initiator, contributed to promoter activity; however, the presence of a consensus initiator in this region was neither necessary nor sufficient for transcription. We concluded from these results that the Sp1 binding site at - 49 substituted for the missing TATA box and played a major role in start site selection at the CAD promoter.
UR - http://www.scopus.com/inward/record.url?scp=0027980124&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0027980124&partnerID=8YFLogxK
M3 - Article
C2 - 7905000
AN - SCOPUS:0027980124
VL - 269
SP - 2252
EP - 2257
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 3
ER -