Purification and characterization of the stage-specific embyronic enhancer-binding protein SSAP-1

Daniel J. Deangelo, Jeffery Defalco, Geoffrey J. Childs

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

We have demonstrated that a highly conserved segment of DNA between positions -288 and -317 (upstream sequence element IV [USE IV]) is largely responsible for the transcriptional activation of the sea urchin H1-β histone gene during the blastula stage of embryogenesis. This sequence is capable of acting as an embryonic enhancer element, activating target genes in a stage-specific manner. Nuclear extracts prepared from developmentally-staged organisms before and after the gene is activated all contain a factor which specifically binds to the enhancer. We have purified a 43-kDa polypeptide which binds to and footprints the USE IV enhancer element. We refer to this protein as stage-specific activator protein 1 (SSAP-1). Early in development before the enhancer is active, SSAP appears as a 43-kDa monomer, but it undergoes a change in its molecular weight beginning at about 12 h postfertilization (early blastula) which precisely parallels the increase in H1-β gene expression. Modified SSAP has an apparent molecular mass of approximately 90 to 100 kDa and contains at least one 43-kDa SSAP polypeptide. Thus, it is the disappearance of the 43-kDa species and the appearance of the 90- to 100-kDa species which coincide with the H1-β gene activation. The correlation between the change in molecular weight of SSAP and the stage-specific activation of H1-β gene expression strongly suggests that this higher-molecular-weight form of SSAP is directly responsible for the blastula stage-specific transcriptional activation of the late H1 gene.

Original languageEnglish (US)
Pages (from-to)1746-1758
Number of pages13
JournalMolecular and Cellular Biology
Volume13
Issue number3
StatePublished - Mar 1993

Fingerprint

Transcription Factor AP-1
Blastula
Carrier Proteins
Transcriptional Activation
Molecular Weight
Genes
Gene Expression
Peptides
Sea Urchins
Histones
Embryonic Development
DNA
Proteins

ASJC Scopus subject areas

  • Cell Biology
  • Genetics
  • Molecular Biology

Cite this

Purification and characterization of the stage-specific embyronic enhancer-binding protein SSAP-1. / Deangelo, Daniel J.; Defalco, Jeffery; Childs, Geoffrey J.

In: Molecular and Cellular Biology, Vol. 13, No. 3, 03.1993, p. 1746-1758.

Research output: Contribution to journalArticle

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