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

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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.

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Authors and Affiliations

Authors

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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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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