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‘See-saw’ expression of microRNA-198 and FSTL1 from a single transcript in wound healing

Nature volume 495, pages 103–106 (2013)Cite this article

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Abstract

Post-transcriptional switches are flexible effectors of dynamic changes in gene expression1. Here we report a new post-transcriptional switch that dictates the spatiotemporal and mutually exclusive expression of two alternative gene products from a single transcript. Expression of primate-specific exonic microRNA-198 (miR-198)2, located in the 3′-untranslated region of follistatin-like 1 (FSTL1)3 messenger RNA, switches to expression of the linked open reading frame of FSTL1 upon wounding in a human ex vivo organ culture system. We show that binding of a KH-type splicing regulatory protein (KSRP, also known as KHSRP) to the primary transcript determines the fate of the transcript and is essential for the processing of miR-198: transforming growth factor-β signalling switches off miR-198 expression by downregulating KSRP, and promotes FSTL1 protein expression. We also show that FSTL1 expression promotes keratinocyte migration, whereas miR-198 expression has the opposite effect by targeting and inhibiting DIAPH1, PLAU and LAMC2. A clear inverse correlation between the expression pattern of FSTL1 (pro-migratory) and miR-198 (anti-migratory) highlights the importance of this regulatory switch in controlling context-specific gene expression to orchestrate wound re-epithelialization. The deleterious effect of failure of this switch is apparent in non-healing chronic diabetic ulcers, in which expression of miR-198 persists, FSTL1 is absent, and keratinocyte migration, re-epithelialization and wound healing all fail to occur.

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Figure 1: Expression of an exonic miRNA or linked open reading frame in context-specific physiological states.
Figure 2: The regulatory switch is impaired in chronic wounds.
Figure 3: KSRP and TGF-β1 regulate the choice between expression of miR-198 or FSTL1.

Accession codes

Primary accessions

Gene Expression Omnibus

Data deposits

Microarray data are deposited in the Gene Expression Omnibus (GEO) under accession numbers GSE37967 and GSE41615.

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Acknowledgements

This work was supported by an A*STAR Investigatorship award to P.S., the Biomedical Research Council of Singapore and the Skin Biology Cluster Platform, A*STAR. We thank T. Kamala, S. Nama, M. Hisyam and C. Vaz for experimental support and B. Knowles for critical reading of the manuscript.

Author information

Author notes

  1. Gopinath M. Sundaram and John E. A. Common: These authors contributed equally to this work.

Authors and Affiliations

  1. Institute of Medical Biology, Agency for Science Technology & Research (A*STAR), 138648, Singapore,

    Gopinath M. Sundaram, John E. A. Common, Felicia E. Gopal, Declan P. Lunny, Vivek Tanavde, E. Birgitte Lane & Prabha Sampath

  2. Jnana Sanjeevini Diabetes Center, Bangalore 560078, India,

    Satyanarayana Srikanta & Krishnaswamy Lakshman

  3. Division of Plastic, Reconstructive & Aesthetic Surgery, National University Health System, 119074, Singapore,

    Thiam C. Lim

  4. Department of Surgery, Yong Loo Lin School of Medicine, National University of Singapore, 119228, Singapore,

    Thiam C. Lim

  5. Bioinformatics Institute, Agency for Science Technology & Research (A*STAR), 138671, Singapore,

    Vivek Tanavde

  6. Department of Pathology, Yong Loo Lin School of Medicine, National University of Singapore, 119074, Singapore,

    E. Birgitte Lane

  7. Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, 117597, Singapore,

    E. Birgitte Lane & Prabha Sampath

Authors
  1. Gopinath M. Sundaram
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Contributions

G.M.S. and J.E.A.C. performed most of the experiments; F.E.G. and D.P.L. helped with immunohistochemistry; K.L., T.C.L. and S.S. assisted in procurement of patient samples; V.T. helped in microarray data analysis; E.B.L. assisted in experimental design and contributed to writing the manuscript; and P.S. designed experiments, supervised this work and wrote the manuscript.

Corresponding author

Correspondence to Prabha Sampath.

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Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-17 and lists of primers a-c. (PDF 1176 kb)

Live cell imaging of scratch wound assay in keratinocytes over-expressing miR-198

Keratinocytes over-expressing miR-198 were plated as a monolayer and subjected to scratch wound assay. Video represents time lapse imaging performed every one hour after wounding for a period of 48 hours. (MOV 4840 kb)

Live cell imaging of scratch wound assay in keratinocytes over-expressing control miRNA

Keratinocytes over-expressing control miRNA were plated as a monolayer and subjected to scratch wound assay. Video represents time lapse imaging performed every one hour after wounding for a period of 48 hours. (MOV 4832 kb)

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Sundaram, G., Common, J., Gopal, F. et al. ‘See-saw’ expression of microRNA-198 and FSTL1 from a single transcript in wound healing. Nature 495, 103–106 (2013). https://doi.org/10.1038/nature11890

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