The bovine DNA polymerase beta promoter: cloning, characterization and comparison with the human core promoter

Gene. 1995 Oct 27;164(2):323-7. doi: 10.1016/0378-1119(95)00498-u.

Abstract

The core promoter of the human DNA polymerase beta (beta Pol)-encoding gene (POL beta) is regulated through cis-elements for the ATF/CREB protein(s), and GC box-binding and initiation-site-binding proteins. The mechanism of promoter regulation has been studied using a nuclear extract transcription system from HeLa cells [Narayan et al., J. Biol. Chem. 269 (1994) 12755-12763]. To study the homologous promoter (ppol beta) in a bovine system, we cloned and characterized the 5'-flanking region of the bovine gene (pol beta). A 15.3-kb fragment of bovine genomic DNA containing the first two exons and 11 kb of 5'-flanking region was isolated from a testis library in bacteriophage lambda EMBL3. S1 nuclease mapping and primer extension analysis of the 5'-end of the pol beta mRNA identified the major transcription start point (tsp), which is located 142-bp 5' of the translational start codon. In transient expression assays using a bovine cell line, analysis of various 5'-deletion mutants demonstrated that a fragment of only 91-bp 5' of the tsp had promoter activity similar to that of a 1.37-kb fragment, so that cis-elements for basal transcription are located within this approx. 100-bp core promoter, as in the human promoter (pPOL beta). Comparison of the core promoters from the bovine and human genes revealed striking similarity, including an almost precise match of the tsp, the ATF/CREB-binding and Sp1-binding sites, and the spacing separating them.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Base Sequence
  • Cattle / genetics*
  • Cell Line
  • Cell Nucleus / metabolism
  • DNA Polymerase I / biosynthesis*
  • DNA Polymerase I / genetics*
  • Gene Expression
  • Gene Expression Regulation, Enzymologic
  • HeLa Cells
  • Hominidae / genetics*
  • Humans
  • Molecular Sequence Data
  • Mutagenesis
  • Promoter Regions, Genetic*
  • Recombinant Proteins / biosynthesis
  • Restriction Mapping
  • Sequence Deletion
  • Sequence Homology, Nucleic Acid
  • Species Specificity
  • Transcription, Genetic
  • Transfection

Substances

  • Recombinant Proteins
  • DNA Polymerase I

Associated data

  • GENBANK/S80341