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Original Article | Open Access

Transcript shortening via alternative polyadenylation promotes gene expression during fracture healing

Deepak Kumar Khajuria1,2, Irena Nowak1,2, Ming Leung3, Vengadeshprabhu Karuppagounder1,2, Yuka Imamura3,4, Christopher C. Norbury5, Fadia Kamal1,2,4, Reyad A. Elbarbary1,2,6,7 ( )
Department of Orthopaedics and Rehabilitation, The Pennsylvania State University College of Medicine, Hershey, PA 17033, USA
Center for Orthopaedic Research and Translational Science (CORTS), The Pennsylvania State University College of Medicine, Hershey, PA 17033, USA
Institute for Personalized Medicine, Penn State College of Medicine, Hershey, PA 17033, USA
Department of Pharmacology, The Pennsylvania State University College of Medicine, Hershey, PA 17033, USA
Department of Microbiology and Immunology, The Pennsylvania State University College of Medicine, Hershey, PA 17033, USA
Department of Biochemistry and Molecular Biology, The Pennsylvania State University College of Medicine, Hershey, PA 17033, USA
Center for RNA Molecular Biology, Pennsylvania State University, University Park, PA 16802, USA
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Abstract

Maturation of the 3′ end of almost all eukaryotic messenger RNAs (mRNAs) requires cleavage and polyadenylation. Most mammalian mRNAs are polyadenylated at different sites within the last exon, generating alternative polyadenylation (APA) isoforms that have the same coding region but distinct 3′ untranslated regions (UTRs). The 3′UTR contains motifs that regulate mRNA metabolism; thus, changing the 3′UTR length via APA can significantly affect gene expression. Endochondral ossification is a central process in bone healing, but the impact of APA on gene expression during this process is unknown. Here, we report the widespread occurrence of APA, which impacts multiple pathways that are known to participate in bone healing. Importantly, the progression of endochondral ossification involves global 3′UTR shortening, which is coupled with an increased abundance of shortened transcripts relative to other transcripts; these results highlight the role of APA in promoting gene expression during endochondral bone formation. Our mechanistic studies of transcripts that undergo APA in the fracture callus revealed an intricate regulatory network in which APA enhances the expression of the collagen, type Ⅰ, alpha 1 (Col1a1) and Col1a2 genes, which encode the 2 subunits of the abundantly expressed protein collagen 1. APA exerts this effect by shortening the 3′UTRs of the Col1a1 and Col1a2 mRNAs, thus removing the binding sites of miR-29a-3p, which would otherwise strongly promote the degradation of both transcripts. Taken together, our study is the first to characterize the crucial roles of APA in regulating the 3′UTR landscape and modulating gene expression during fracture healing.

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Bone Research
Volume 11,
December 2023
Article number: 5
DOI: 10.1038/s41413-022-00236-7
Cite this article:
Khajuria DK, Nowak I, Leung M, et al. Transcript shortening via alternative polyadenylation promotes gene expression during fracture healing. Bone Research, 2023, 11: 5. https://doi.org/10.1038/s41413-022-00236-7

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Received: 15 June 2022
Revised: 15 September 2022
Accepted: 12 October 2022
Published: 03 January 2023
© The Author(s) 2023

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