Complex Transcriptional Profiles of the PPP1R12A Gene in Cells of the Circulatory System as Revealed by In Silico Analysis and Reverse Transcription PCR

Saldanha, Paulo André; Bolanle, Israel Olapeju; Palmer, Timothy Martin; Nikitenko, Leonid Leonidovich; Rivero, Francisco

Abstract

The myosin light chain phosphatase target subunit 1 (MYPT1), encoded by the PPP1R12A gene, is a key component of the myosin light chain phosphatase (MLCP) protein complex. MYPT1 isoforms have been described as products of the cassette-type alternative splicing of exons E13, E14, E22, and E24. Through in silico analysis of the publicly available EST and mRNA databases, we established that PPP1R12A contains 32 exons (6 more than the 26 previously reported), of which 29 are used in 11 protein-coding transcripts. An in silico analysis of publicly available RNAseq data combined with validation by reverse transcription (RT)-PCR allowed us to determine the relative abundance of each transcript in three cell types of the circulatory system where MYPT1 plays important roles: human umbilical vein endothelial cells (HUVEC), human saphenous vein smooth muscle cells (HSVSMC), and platelets. All three cell types express up to 10 transcripts at variable frequencies. HUVECs and HSVSMCs predominantly express the full-length variant (58.3% and 64.3%, respectively) followed by the variant skipping E13 (33.7% and 23.1%, respectively), whereas in platelets the predominant variants are those skipping E14 (51.4%) and E13 (19.9%), followed by the full-length variant (14.4%). Variants including E24 account for 5.4% of transcripts in platelets but are rare (<1%) in HUVECs and HSVSMCs. Complex transcriptional profiles were also found across organs using in silico analysis of RNAseq data from the GTEx project. Our findings provide a platform for future studies investigating the specific (patho)physiological roles of understudied MYPT1 isoforms.

Citation

Saldanha, P. A., Bolanle, I. O., Palmer, T. M., Nikitenko, L. L., & Rivero, F. (2022). Complex Transcriptional Profiles of the PPP1R12A Gene in Cells of the Circulatory System as Revealed by In Silico Analysis and Reverse Transcription PCR. Cells, 11(15), Article 2315. https://doi.org/10.3390/cells11152315

• Journal Article Type: Article
• Acceptance Date: Jul 25, 2022
• Online Publication Date: Jul 27, 2022
• Publication Date: Aug 1, 2022
• Deposit Date: Jul 27, 2022
• Publicly Available Date: Aug 1, 2022
• Journal: Cells
• Electronic ISSN: 2073-4409
• Publisher: MDPI
• Peer Reviewed: Peer Reviewed
• Volume: 11
• Issue: 15
• Article Number: 2315
• DOI: https://doi.org/10.3390/cells11152315
• Keywords: MYPT1; PPP1R12A; Alternative splicing; Promoter; Terminator; HUVEC; HSVSMC; Platelet
• Public URL: https://hull-repository.worktribe.com/output/4040853