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PMID: 8887666 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Recruitment of the tinman homolog Nkx-2.5 by serum response factor activates cardiac alpha-actin gene transcription.

Molecular and cellular biology ·Vol. 16 ·No. 11 ·1996-11-00 ·Pages 6372-84

Chen CY, Schwartz RJ

Abstract

We recently showed that the cardiogenic homeodomain factor Nkx-2.5 served as a positive acting accessory factor for serum response factor (SRF) and that together they provided strong transcriptional activation of the cardiac alpha-actin promoter, depending upon intact serum response elements (SREs) (C. Y. Chen, J. Croissant, M. Majesky, S. Topouz, T. McQuinn, M. J. Frankovsky, and R. J. Schwartz, Dev. Genet. 19:119-130, 1996). As shown here, Nkx-2.5 and SRF collaborated to activate the endogenous murine cardiac alpha-actin gene in 10T1/2 fibroblasts by a mechanism in which SRF recruited Nkx-2.5 to the alpha-actin promoter. Activation of a truncated promoter consisting of the proximal alpha-actin SRE1 occurred even when Nkx-2.5 DNA-binding activity was blocked by a point mutation in the third helix of its homeodomain. Investigation of protein-protein interactions showed that Nkx-2.5 was bound to SRF in the absence of DNA in soluble protein complexes retrieved from cardiac myocyte nuclei but could also be detected in coassociated binding complexes on the proximal SRE1. Recruitment of Nkx-2.5 to an SRE depended upon SRF DNA-binding activity and was blocked by the dominant negative SRFpm1 mutant, which allowed for dimerization of SRF monomers but prevented DNA binding. Interactive regions shared by Nkx-2.5 and SRF were mapped to N-terminal/helix I and helix II/helix III regions of the Nkx-2.5 homeodomain and to the N-terminal extension of the MADS box. Our study suggests that physical association between Nkx-2.5 and SRF is one way that cardiac specified genes are activated in cardiac cell lineages.

MeSH Terms
Actins/biosynthesis Animals Base Sequence Binding Sites Cell Line Cells, Cultured Chickens DNA-Binding Proteins/metabolism Homeobox Protein Nkx-2.5 Homeodomain Proteins/metabolism Humans Mice Mice, Inbred C3H Myocardium/metabolism Nuclear Proteins/metabolism Polymerase Chain Reaction Promoter Regions, Genetic Protein Biosynthesis Recombinant Fusion Proteins/biosynthesis,metabolism Serum Response Factor TATA Box Transcription Factors/metabolism Transcription, Genetic Transcriptional Activation Transfection Xenopus Proteins
Chemicals
Actins DNA-Binding Proteins Homeobox Protein Nkx-2.5 Homeodomain Proteins NKX2-5 protein, human Nkx2-5 protein, mouse Nuclear Proteins Recombinant Fusion Proteins Serum Response Factor Transcription Factors Xenopus Proteins
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Chen C Y
Department of Cell Biology, Baylor College of Medicine, Houston, Texas 77030, USA.
Schwartz R J
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.16.11.6372
Published
1996-11-00
Pages
6372-84
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC231639
Subset
IM
Grants
NHLBI NIH HHS · P01 HL49953 · United States
NHLBI NIH HHS · R01 HL50422 · United States
External Links
PubMed source DOI: 10.1128/MCB.16.11.6372 PMC Full Text View journal details
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