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Serum response factor regulates a muscle-specific microRNA that _targets Hand2 during cardiogenesis

Nature volume 436, pages 214–220 (2005)Cite this article

Abstract

Gradients of signalling and transcription factors govern many aspects of embryogenesis, highlighting the need for spatiotemporal control of regulatory protein levels. MicroRNAs are phylogenetically conserved small RNAs that regulate the translation of _target messenger RNAs, providing a mechanism for protein dose regulation. Here we show that microRNA-1-1 (miR-1-1) and miR-1-2 are specifically expressed in cardiac and skeletal muscle precursor cells. We found that the miR-1 genes are direct transcriptional _targets of muscle differentiation regulators including serum response factor, MyoD and Mef2. Correspondingly, excess miR-1 in the developing heart leads to a decreased pool of proliferating ventricular cardiomyocytes. Using a new algorithm for microRNA _target identification that incorporates features of RNA structure and _target accessibility, we show that Hand2, a transcription factor that promotes ventricular cardiomyocyte expansion, is a _target of miR-1. This work suggests that miR-1 genes titrate the effects of critical cardiac regulatory proteins to control the balance between differentiation and proliferation during cardiogenesis.

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Figure 1: miR-1 s are highly conserved and are cardiac- and skeletal-muscle-specific.
Figure 2: SRF, Mef2 and MyoD directly regulate embryonic expression of miR-1.
Figure 3: miR-1 regulates pool of proliferating ventricular cardiomyocytes and ventricular expansion.
Figure 4: Prediction and validation of miR-1 _targets.

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Acknowledgements

We wish to thank K. Ivey for critical discussions and for preparation of figures; members of the Srivastava laboratory for helpful discussions; J. McAnally for generation of transgenic mice; E. N. Olson for plasmids; and R. Misra and R. Balza for providing SRF-null embryonic heart cDNA. D. S. was supported by grants from NHLB/NIH, the March of Dimes Birth Defects Foundation and the American Heart Association.

Author information

Author notes

  1. Yong Zhao, Eva Samal & Deepak Srivastava

    Present address: Gladstone Institute of Cardiovascular Disease and Department of Pediatrics, University of California San Francisco, 1650 Owens Street, San Francisco, California, 94158, USA

Authors and Affiliations

  1. Department of Pediatrics (Cardiology),

    Yong Zhao, Eva Samal & Deepak Srivastava

  2. Molecular Biology, University of Texas Southwestern Medical Center and Children's Medical Center Dallas, 6000 Harry Hines Boulevard, Texas, 75390-9148, Dallas, USA

    Yong Zhao, Eva Samal & Deepak Srivastava

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Correspondence to Deepak Srivastava.

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Competing interests

MicroRNA 1 sequences have been deposited in Genbank under the following accession numbers: DQ066648 and DQ 066649 (Pan troglodytes pre-microRNA-1-1 and 1-2), DQ066650 (rat pre-microRNA-1), DQ066651 (Danio rerio pre-microRNA-1) and DQ066652 (Xenopus tropicalis pre-microRNA-1). Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.

Supplementary information

Supplementary Figure S1

Processed miR-1 is 21bp in length. This figure shows that endogenous mature form of miR-1 in skeletal muscle and heart is 21bp. (DOC 362 kb)

Supplementary Figure S2

Alignment of known microRNA _targets from different species. This figure shows that virtually all low δG sites are completely conserved, whereas high δG sites have variances in the sequence matching the critical 5' region of the miRNA. (DOC 100 kb)

Supplementary Figure S3

Conserved miR-1 _target sites in Hand2, TB4 and IGF1 3' UTRs. This figure shows sequence complementarity between miR-1 and 3' UTRs of Hand2, TB4 and IGF1 . (DOC 23 kb)

Supplementary Figure S4

miR-1 _target validation. This figure shows successful expression of mutant miR-1. Wild type or mutant miR-1s can repress the corresponding 3'UTRs or their mutant forms, respectively. (DOC 421 kb)

Supplementary Figure S5

Transgenic overexpression of αMHC-miR-1. This figure shows that miR-1 can be successfully highly expressed in heart in vivo under control of an alpha-MHC promoter. (DOC 247 kb)

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Zhao, Y., Samal, E. & Srivastava, D. Serum response factor regulates a muscle-specific microRNA that _targets Hand2 during cardiogenesis. Nature 436, 214–220 (2005). https://doi.org/10.1038/nature03817

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