An alternative Pit-1 RNA splicing product reveals modular binding and nonmodular transcriptional activities of the POU-specific domain

Jeffrey W. Voss, Laura Wilson, Simon Rhodes, Michael G. Rosenfeld

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

We have identified a form of the pituitary-specific POU protein Pit-1 that results from deletion of the POU-specific (POUs) domain by alternative RNA splicing. This natural variant of Pit-1 (called Δ 4Pit-1) has revealed several aspects of the function of the POUs domain. The Δ 4Pit-1 protein was characterized using a Δ 4Pit-1-specific antiserum. Further, selection assays of random oligonucleotide pools identified binding site preferences for both wild type and Δ 4Pit-1. Methylation interference, copper phenanthrolene, and missing contact analyses were used to compare the binding characteristics of the two forms of Pit-1 on a selected site. DNA binding affinity assays on several DNA elements revealed that the POUs domain contains a modular DNA binding activity affecting the DNA binding affinity of the entire POU domain on some, but not on other, DNA sites. Functional analysis on such DNA elements has revealed that the POUs domain is an essential, but nonmodular, component of the Pit-1 trans-activation domain dependent on its natural context within the Pit-1 protein.

Original languageEnglish (US)
Pages (from-to)1551-1560
Number of pages10
JournalMolecular Endocrinology
Volume7
Issue number12
DOIs
StatePublished - 1993
Externally publishedYes

Fingerprint

RNA Splicing
DNA
POU Domain Factors
Alternative Splicing
Oligonucleotides
Methylation
Immune Sera
Copper
Proteins
Binding Sites

ASJC Scopus subject areas

  • Molecular Biology
  • Endocrinology

Cite this

An alternative Pit-1 RNA splicing product reveals modular binding and nonmodular transcriptional activities of the POU-specific domain. / Voss, Jeffrey W.; Wilson, Laura; Rhodes, Simon; Rosenfeld, Michael G.

In: Molecular Endocrinology, Vol. 7, No. 12, 1993, p. 1551-1560.

Research output: Contribution to journalArticle

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