Advertisement
JOURNAL ARTICLES
An HF-1a/HF-1b/MEF-2 combinatorial element confers cardiac ventricular specificity and established an anterior-posterior gradient of expression
R.S. Ross, S. Navankasattusas, R.P. Harvey, K.R. Chien
Development 1996 122: 1799-1809;

Summary

The molecular determinants that direct gene expression to the ventricles of the heart are for the most part unknown. Additionally, little data is available on how the anterior/posterior axis of the heart tube is determined and whether the left and right atrial and ventricular chambers are assigned as part of this process. Utilizing myosin light chain-2 ventricular promoter/beta-galactosidase reporter transgenes, we have determined the minimal cis-acting sequences required for ventricular-specific gene expression. In multiple independent transgenic mouse lines, we found that both a 250 base pair myosin light chain-2 ventricular promoter fragment, as well as a dimerized 28 bp sub-element (HF-1) containing binding sites for HF1a and HF1b/MEF2 factors, directed ventricular-specific reporter expression from as early as the endogenous gene, at day 7.5-8.0 post coitum. While the endogenous gene is expressed uniformly throughout both ventricles, the transgenes were expressed in a right ventricular/conotruncal dominant fashion, suggesting that they contain only a subset of the elements which respond to positional information in the developing heart tube. Expression of the transgene was cell autonomous and its temporospatial characteristics not affected by mouse strain/methylation state of the genome. To determine whether ventricular-specific expression of the transgene was dependent upon regulatory genes required for correct ventricular differentiation, the 250 base pair transgene was bred into both retinoid X receptoralpha and Nkx2-5 null backgrounds. The transgene was expressed in both mutant backgrounds, despite the absence of endogenous myosin light chain-2 ventricular transcript in Nkx2-5 null embryos. Ventricular specification, as judged by transgene expression, appeared to occur normally in both mutants. Thus, the HF-1 element, directs chamber-specific transcription of a transgene reporter independently of retinoid X receptoralpha and Nkx2-5, and defines a minimal combinatorial pathway for ventricular chamber gene expression. The patterned expression of this transgene may provide a model system in which to investigate the cues that dictate anterior-posterior (right ventricle/left ventricle) gradients during mammalian heart development.

Reference

    1. Chien K. R.,
    2. Zhu H.,
    3. Knowlton K.U.,
    4. Miller-Hance W.,
    5. van-Bilsen M.,
    6. O'Brien T.X.,
    7. Evans S. M.
    (1993) Transcriptional regulation during cardiac growth and development. Annu. Rev. Physiol 55, 77–95
    OpenUrlCrossRefPubMedWeb of Science
    1. Cserjesi P.,
    2. Olson E. N.
    (1991) Myogenin induces the myocyte-specific enhancer binding factor MEF-2 independently of other muscle-specific gene products. Mol. Cell. Biol 11, 4854–4862
    OpenUrlAbstract/FREE Full Text
    1. Donoghue M. J.,
    2. Merlie J. P.,
    3. Rosenthal N.,
    4. Sanes J. R.
    (1991) Rostrocaudal gradient of transgene expression in adult skeletal muscle. Proc. Natl. Acad. Sci. USA 88, 5847–5851
    OpenUrlAbstract/FREE Full Text
    1. Engler P.,
    2. Haasch D.,
    3. Pinkert C. A.,
    4. Doglio L.,
    5. Glymour M.,
    6. Brinster R.,
    7. Storb U.
    (1991) A strain-specific modifier on mouse chromosome 4 controls the methylation of independent transgene loci. Cell 65, 939–947
    OpenUrlCrossRefPubMedWeb of Science
    1. Evans S. M.,
    2. Tai L. J.,
    3. Tan V. P.,
    4. Newton C. B.,
    5. Chien K. R.
    (1994) Heterokaryons of cardiac myocytes and fibroblasts reveal the lack of dominance of the cardiac muscle phenotype. Mol. Cell Biol 14, 4269–4279
    OpenUrlAbstract/FREE Full Text
    1. Grieshammer U.,
    2. Sassoon D.,
    3. Rosenthal N.
    (1992) A transgene target for positional regulators marks early rostrocaudal specification of myogenic lineages. Cell 69, 79–93
    OpenUrlCrossRefPubMedWeb of Science
    1. Henderson S. A.,
    2. Spencer M.,
    3. Sen A.,
    4. Kumar C.,
    5. Siddiqui M. A.,
    6. Chien K. R.
    (1989) Structure, organization, and expression of the rat cardiac myosin light chain-2 gene. Identification of a 250-base pair fragment which confers cardiac-specific expression. J. Biol. Chem 264, 18142–18148
    OpenUrlAbstract/FREE Full Text
    1. Hoyle C.,
    2. Brown N. A.,
    3. Wolpert L.
    (1992) Development of left/right handedness in the chick heart. Development 115, 1071–1078
    OpenUrlAbstract
    1. Hunter J. J.,
    2. Tanaka N.,
    3. Rockman H. A.,
    4. Ross Jr..,
    5. Chien K. R.
    (1995) Ventricular expression of a MLC-2v- Ras fusion gene induces cardiac hypertrophy and selective diastolic dysfunction in transgenic mice. J. Biol. Chem 270, 23173–23178
    OpenUrlAbstract/FREE Full Text
    1. Kelly R.,
    2. Alonso S.,
    3. Tajbakhsh S.,
    4. Cossu G.,
    5. Buckingham M.
    (1995) Myosin light chain 3F regulatory sequences confer regionalized cardiac and skeletal muscle expression in transgenic mice. J. Cell Biol 129, 383–396
    OpenUrlAbstract/FREE Full Text
    1. Lee K. J.,
    2. Ross R. S.,
    3. Rockman H. A.,
    4. Harris A. N.,
    5. O'Brien T. X.,
    6. van Bilsen M.,
    7. Shubeita H. E.,
    8. Kandolf R.,
    9. Brem G.,
    10. Price J.,
    11. Chien K. R.
    (1992) Myosin light chain-2 luciferase transgenic mice reveal distinct regulatory. J. Biol. Chem 267, 15875–15885
    OpenUrlAbstract/FREE Full Text
    1. Lee K. J.,
    2. Hickey R.,
    3. Zhu H.,
    4. Chien K. R.
    (1994) Positive regulatory elements (HF-1a and HF-1b) and a novel negative regulatory element (HF-3) mediate ventricular muscle-specific expression of myosin light-chain 2-luciferase fusion genes in transgenic mice. Mol. Cell Biol 14, 1220–1229
    OpenUrlAbstract/FREE Full Text
    1. Levin M.,
    2. Johnson R. L.,
    3. Stern C. D.,
    4. Kuehn M.,
    5. Tabin C.
    (1995) A molecular pathway determining left-right asymmetry in chick embryogenesis. Cell 82, 803–814
    OpenUrlCrossRefPubMedWeb of Science
    1. Lilly B.,
    2. Zhao B.,
    3. Ranganayakulu G.,
    4. Paterson B. M.,
    5. Schulz R. A.,
    6. Olson E. N.
    (1995) Requirement of MADS domain transcription factor D-MEF2 for muscle formation in Drosophila. Science 267, 688–93
    OpenUrlAbstract/FREE Full Text
    1. Lyons G. E.,
    2. Schiaffino S.,
    3. Sassoon D.,
    4. Barton P.,
    5. Buckingham M.
    (1990) Developmental regulation of myosin gene expression in mouse cardiac muscle. J. Cell Biol 111, 2427–2436
    OpenUrlAbstract/FREE Full Text
    1. Lyons I.,
    2. Parsons L. M.,
    3. Hartley L.,
    4. Li R.,
    5. Andrews J. E.,
    6. Robb L.,
    7. Harvey R. P.
    (1995) Myogenic and morphogenic defects in the heart tubes of murine embryos lacking the homeo box gene Nkx2-5. Genes Dev 9, 1654–1666
    OpenUrlAbstract/FREE Full Text
    1. Molkentin J. D.,
    2. Brogan R. S.,
    3. Jobe S. M.,
    4. Markham B. E.
    (1993) Expression of the alpha-myosin heavy chain gene in the heart is regulated in part by an E-box-dependent mechanism. J. Biol. Chem 268, 2602–2609
    OpenUrlAbstract/FREE Full Text
    1. Navankasattusas S.,
    2. Zhu H.,
    3. Garcia A. V.,
    4. Evans S. M.,
    5. Chien K. R.
    (1992) A ubiquitous factor (HF-1a) and a distinct muscle factor (HF-1b/MEF-2) form an E-box-independent pathway for cardiac muscle gene expression. Mol. Cell. Biol 12, 1469–1479
    OpenUrlAbstract/FREE Full Text
    1. Navankasattusas S.,
    2. Sawadogo M.,
    3. van Bilsen M.,
    4. Dang C. V.,
    5. Chien K. R.
    (1994) The basic helix-loop-helix protein upstream stimulating factor regulates the cardiac ventricular myosin light-chain 2 gene via independent cis regulatory elements. Mol. Cell Biol 14, 7331–7339
    OpenUrlAbstract/FREE Full Text
    1. Nordeen S. K.
    (1988) Luciferase reporter gene vectors for analysis of promoters and enhancers. BioTechniques 6, 454–457
    OpenUrlPubMedWeb of Science
    1. O'Brien T. X.,
    2. Lee K. J.,
    3. Chien K. R.
    (1993) Positional specification of ventricular myosin light chain 2 expression in the primitive murine heart tube. Proc. Natl. Acad. Sci. USA 90, 5157–5161
    OpenUrlAbstract/FREE Full Text
    1. Ozer J.,
    2. Faber M.,
    3. Chalkley R.,
    4. Sealy L.
    (1990) Isolation and characterization of a cDNA clone for the CCAAT transcription factor EFIA reveals a novel structural motif. J. Biol. Chem 265, 22143–22152
    OpenUrlAbstract/FREE Full Text
    1. Sanes J. R.,
    2. Rubenstein J. L.,
    3. Nicolas J. F.
    (1986) Use of a recombinant retrovirus to study post-implantation cell lineage in mouse embryos. EMBO J 5, 3133–3142
    OpenUrlPubMedWeb of Science
    1. Shubeita H. E.,
    2. Thorburn J.,
    3. Chien K. R.
    (1992) Microinjection of antibodies and expression vectors into living myocardial cells. Development of a novel approach to identify candidate genes that regulate cardiac growth and hypertrophy. Circulation 85, 2236–2246
    OpenUrlAbstract/FREE Full Text
    1. Sucov H. M.,
    2. Dyson E.,
    3. Gumeringer C. L.,
    4. Price J.,
    5. Chien K. R.,
    6. Evans R. M.
    (1994) RXR alpha mutant mice establish a genetic basis for vitamin A signaling in heart morphogenesis. Genes Dev 8, 1007–1018
    OpenUrlAbstract/FREE Full Text
    1. Yang J. T.,
    2. Rayburn H.,
    3. Hynes R. O.
    (1995) Cell adhesion events mediated by alpha 4 integrins are essential in placental and cardiac development. Development 121, 549–560
    OpenUrlAbstract
    1. Yu Y. T.,
    2. Breitbart R. E.,
    3. Smoot L. B.,
    4. Lee Y.,
    5. Mahdavi V.,
    6. Nadal-Ginard B.
    (1992) Human myocyte-specific enhancer factor 2 comprises a group of tissue-restricted MADS box transcription factors. Genes Dev 6, 1783–1798
    OpenUrlAbstract/FREE Full Text
    1. Zeller R.,
    2. Bloch K. D.,
    3. Williams B. S.,
    4. Arceci R. J.,
    5. Seidman C. E.
    (1987) Localized expression of the atrial natriuretic factor gene during cardiac embryogenesis. Genes Dev 1, 693–698
    OpenUrlAbstract/FREE Full Text
    1. Zhu H.,
    2. Garcia A. V.,
    3. Ross R. S.,
    4. Evans S. M.,
    5. Chien K. R.
    (1991) A conserved 28-base-pair element (HF-1) in the rat cardiac myosin light-chain-2 gene confers cardiac-specific and alpha-adrenergic-inducible expression in cultured neonatal rat myocardial cells. Mol. Cell. Biol 11, 2273–2281
    OpenUrlAbstract/FREE Full Text
    1. Zou Y.,
    2. Chien K. R.
    (1995) EFIA/YB-1 is a component of cardiac HF-1A binding activity and positively regulates transcription of the myosin light-chain 2v gene. Mol. Cell Biol 15, 2972–2982
    OpenUrlAbstract/FREE Full Text
Back to top

 Download PDF

Email
JOURNAL ARTICLES
An HF-1a/HF-1b/MEF-2 combinatorial element confers cardiac ventricular specificity and established an anterior-posterior gradient of expression
R.S. Ross, S. Navankasattusas, R.P. Harvey, K.R. Chien
Development 1996 122: 1799-1809;
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
Citation Tools
Alerts

Article navigation

Other journals from The Company of Biologists

Advertisement

Kathryn Virginia Anderson (1952-2020)

Developmental geneticist Kathryn Anderson passed away at home on 30 November 2020. Tamara Caspary, a former postdoc and friend, remembers Kathryn and her remarkable contribution to developmental biology.

Zooming into 2021

In a new Editorial, Editor-in-Chief James Briscoe and Executive Editor Katherine Brown reflect on the triumphs and tribulations of the last 12 months, and look towards a hopefully calmer and more predictable year.

Read & Publish participation extends worldwide

Over 60 institutions in 12 countries are now participating in our Read & Publish initiative. Here, James Briscoe explains what this means for his institution, The Francis Crick Institute. Find out more and view our full list of participating institutions.

Upcoming special issues

Imaging Development, Stem Cells and Regeneration
Submission deadline: 30 March 2021
Publication: mid-2021

The Immune System in Development and Regeneration
Guest editors: Florent Ginhoux and Paul Martin
Submission deadline: 1 September 2021
Publication: Spring 2022

Both special issues welcome Review articles as well as Research articles, and will be widely promoted online and at key global conferences.

Development presents...

Our successful webinar series continues into 2021, with early-career researchers presenting their papers and a chance to virtually network with the developmental biology community afterwards. Sign up to join our next session:

10 February
Time: 13:00 (GMT)
Chaired by: preLights