Faculty Collaboration Database

Start site selection at the TATA-less carbamoyl-phosphate synthase (glutamine-hydrolyzing)/aspartate carbamoyltransferase/dihydroorotase promoter. J Biol Chem 1994 Jan 21;269(3):2252-7

Date

01/21/1994

Pubmed ID

7905000

Scopus ID

2-s2.0-0027980124 (requires institutional sign-in at Scopus site)   74 Citations

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.

Author List

Kollmar R, Sukow KA, Sponagle SK, Farnham PJ

Author

Stephen K. Sponagle MD Assistant Professor in the Psychiatry and Behavioral Medicine department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

3T3 Cells
Animals
Aspartate Carbamoyltransferase
Base Sequence
Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing)
Cell Line
Cell Nucleus
Consensus Sequence
Cricetinae
DNA
Dihydroorotase
Liver
Male
Mesocricetus
Mice
Molecular Sequence Data
Multienzyme Complexes
Mutagenesis, Insertional
Mutagenesis, Site-Directed
Oligodeoxyribonucleotides
Plasmids
Promoter Regions, Genetic
Rats
Rats, Sprague-Dawley
Restriction Mapping
Sequence Homology, Nucleic Acid
T-Lymphocytes
TATA Box
Transfection