Binding of a 100-kDa ubiquitous factor to the human prolactin promoter is required for its basal and hormone-regulated activity

Eur J Biochem. 1992 Nov 15;210(1):53-8. doi: 10.1111/j.1432-1033.1992.tb17389.x.

Abstract

cAMP strongly stimulates the activity of the human prolactin (hPRL) promoter. We have previously shown that two types of cis-element are required for this cAMP regulation; binding sites for the pituitary-specific factor Pit-1, and the sequence spanning nucleotides -115 to -85 (named sequence A). Sequence A contains the TGACG motif found in the consensus sequence of the cAMP-responsive element (CRE). In this study, we show that a mutation in the TGACG motif of sequence A strongly reduces not only the cAMP regulation but also the Ca2+ regulation and basal activity of the hPRL promoter. Furthermore, gel-shift assays indicate that the mutation prevents binding of a ubiquitous factor which is not the CRE-binding protein. Southwestern experiments suggest that this ubiquitous factor's molecular mass is approximately 100 kDa. We conclude that binding of a 100-kDa ubiquitous factor to sequence A is required for full basal and hormonal regulation of hPRL-promoter activity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Blotting, Southern
  • Blotting, Western
  • Cells, Cultured
  • Cyclic AMP / metabolism
  • Humans
  • Molecular Sequence Data
  • Mutation
  • Oligonucleotides
  • Plasmids
  • Prolactin / genetics*
  • Prolactin / metabolism
  • Promoter Regions, Genetic*
  • Trans-Activators / metabolism*

Substances

  • Oligonucleotides
  • Trans-Activators
  • Prolactin
  • Cyclic AMP

Associated data

  • GENBANK/M85175
  • GENBANK/M85176
  • GENBANK/M85177
  • GENBANK/X00368
  • GENBANK/X65179
  • GENBANK/X65180
  • GENBANK/X65181
  • GENBANK/X68289
  • GENBANK/X68290
  • GENBANK/X68291