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Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
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Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
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RBPMS and RBPMS2 Cooperate to Safeguard Cardiac Splicing.

Tongbin Wu1, Zeyu Chen2, Chao Gao1

  • 1Department of Biomedical Research and Translational Medicine, Masonic Medical Research Institute, Utica, NY (T.W., C.G., E.V.S.).

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RNA-binding protein with multiple splicing (RBPMS) and RBPMS2 are crucial for heart development. Their combined action ensures proper splicing of cardiac genes, preventing embryonic lethality and sarcomere defects.

Keywords:
RNA splicingRNA splicing factorsalternative splicingcardiomyopathiesmyocytes, cardiacsarcomeres

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Area of Science:

  • Cardiovascular Biology
  • Molecular Genetics
  • Gene Regulation

Background:

  • Mutations in cardiac splicing factors (SFs) lead to cardiomyopathy and congenital heart disease.
  • Cardiac SFs cooperatively regulate essential cardiac gene splicing, but the functional importance of this collaboration is unclear.
  • RBPMS and RBPMS2 are SFs involved in heart development with similar in vitro splicing activities, but their in vivo cooperation is unknown.

Purpose of the Study:

  • To investigate the in vivo cooperation of RBPMS and RBPMS2 in cardiac development.
  • To determine the functional consequences of RBPMS and RBPMS2 loss in cardiomyocytes.
  • To elucidate the molecular mechanisms underlying their collaborative splicing regulation.

Main Methods:

  • Generated and analyzed cardiomyocyte-specific single and double knockout mice for RBPMS and RBPMS2.
  • Performed RNA sequencing to assess gene expression and splicing changes.
  • Utilized in silico analyses and minigene splicing assays to dissect regulatory mechanisms.

Main Results:

  • Double knockout mice lacking both RBPMS and RBPMS2 died before embryonic day 13.5 with sarcomere disarray.
  • Single knockout mice survived to adulthood with normal sarcomere assembly, indicating overlapping functions.
  • Defective sarcomere assembly in double knockouts resulted from widespread mis-splicing of cardiac contraction genes.
  • RBPMS and RBPMS2 collectively promote cardiac splicing and repress non-cardiac splicing, with binding location dictating their role as activators or repressors.

Conclusions:

  • RBPMS and RBPMS2 collaborate to maintain the splicing of genes critical for cardiac contraction.
  • SF collaboration is vital for preserving the cardiac splicing signature.
  • Understanding SF cooperation is essential for developing therapeutic strategies targeting SF activity.