Mechanisms of MARCKS gene activation during Xenopus development

Yi Shi, Stephen K. Sullivan, Diana M. Pitterle, Elizabeth A. Kennington, Jonathan M. Graff, Perry J. Blackshear

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

The myristoylated alanine-rich protein kinase C substrate (MARCKS) is a high affinity cellular substrate for protein kinase C. The MARCKS gene is under multiple modes of transcriptional control, including cytokine-and transformation-dependent, cell-specific, and developmental regulation. This study evaluated the transcriptional control of MARCKS gene expression during early development of Xenopus laevis. Xenopus MARCKS was highly conserved with its mammalian and avian homologues; its mRNA and protein were abundant in the maternal pool and increased after the mid-blastula transition (MBT). The Xenopus MARCKS gene was similar to those of other species, except that a second intron interrupted the 5'- untranslated region. By transiently transfecting XTC-2 cells and microinjecting Xenopus embryos with reporter gene constructs containing serial deletions of 5'-flanking MARCKS sequences, we identified a 124-base pair minimal promoter that was critical for promoter activity. Developmental gel shift assays revealed that a CBF/NF-Y/CP-1-like factor and an Sp1-like factor bound to this region in a manner correlating with the onset of Xenopus MARCKS transcription at MBT. Mutations in the promoter that abolished binding of these two factors also completely inhibited transcriptional activation of the MARCKS gene at MBT. The binding sites for these two factors are highly conserved in the human and mouse MARCKS promoters, suggesting that these elements might also regulate MARCKS transcription in other species. These studies not only increase our knowledge of the transcriptional regulation of the MARCKS genes but also have implications for the mechanisms responsible for zygotic activation of the Xenopus genome at MBT.

Original languageEnglish (US)
Pages (from-to)29290-29300
Number of pages11
JournalJournal of Biological Chemistry
Volume272
Issue number46
DOIs
StatePublished - Nov 14 1997

Fingerprint

Xenopus
Alanine
Protein Kinase C
Transcriptional Activation
Genes
Chemical activation
Substrates
Blastula
Transcription
myristoylated alanine-rich C kinase substrate
5' Untranslated Regions
Xenopus laevis
Reporter Genes
Gene expression
Base Pairing
Introns
Assays
Embryonic Structures
Gels
Binding Sites

ASJC Scopus subject areas

  • Biochemistry

Cite this

Shi, Y., Sullivan, S. K., Pitterle, D. M., Kennington, E. A., Graff, J. M., & Blackshear, P. J. (1997). Mechanisms of MARCKS gene activation during Xenopus development. Journal of Biological Chemistry, 272(46), 29290-29300. https://doi.org/10.1074/jbc.272.46.29290

Mechanisms of MARCKS gene activation during Xenopus development. / Shi, Yi; Sullivan, Stephen K.; Pitterle, Diana M.; Kennington, Elizabeth A.; Graff, Jonathan M.; Blackshear, Perry J.

In: Journal of Biological Chemistry, Vol. 272, No. 46, 14.11.1997, p. 29290-29300.

Research output: Contribution to journalArticle

Shi, Y, Sullivan, SK, Pitterle, DM, Kennington, EA, Graff, JM & Blackshear, PJ 1997, 'Mechanisms of MARCKS gene activation during Xenopus development', Journal of Biological Chemistry, vol. 272, no. 46, pp. 29290-29300. https://doi.org/10.1074/jbc.272.46.29290
Shi Y, Sullivan SK, Pitterle DM, Kennington EA, Graff JM, Blackshear PJ. Mechanisms of MARCKS gene activation during Xenopus development. Journal of Biological Chemistry. 1997 Nov 14;272(46):29290-29300. https://doi.org/10.1074/jbc.272.46.29290
Shi, Yi ; Sullivan, Stephen K. ; Pitterle, Diana M. ; Kennington, Elizabeth A. ; Graff, Jonathan M. ; Blackshear, Perry J. / Mechanisms of MARCKS gene activation during Xenopus development. In: Journal of Biological Chemistry. 1997 ; Vol. 272, No. 46. pp. 29290-29300.
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