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The Prkar1b gene encodes regulatory type 1 beta subunit of protein kinase A. Here we report that mouse R1β gene produces three alternative splice variants (designated as mR1β1, mR1β2 and mR1β3) that have different N-terminal protein structures. New splice variants were identified using combinatorial approach of bioinformatics pipeline involving online available tools and databases, and molecular biology techniques involving RT-PCR, semi-nested PCR and sequencing. Except mR1β3, which was not detected by RT-PCR in brain and muscle tissues of 3day old mice, all three spliced isoforms were found to be ubiquitously expressed in tissues and postnatal developmental stages examined. The presence of different N-termini in isoforms may be important for unique docking interactions with A kinase anchoring proteins., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Prkaca gene of mouse encodes for a cAMP dependent protein kinase catalytic alpha subunit. PKA occurs naturally as a 4-membered structure having two regulatory (R) and two catalytic (C) subunits each encoded by separate gene. Alternatively spliced two transcript variants are known for the Prkaca gene, which encode for two isoforms of PKA C-subunits, namely Cα1 and Cα2. These isoforms arise as a result of alternative splicing of the first coding exon with the internal exons. We have identified a new transcript variant using combinatorial approach of bioinformatics and molecular biology techniques involving RT-PCR, semi-nested PCR and sequencing. The new transcript variant encoding Cα3 isoform has N-terminus that differs from Cα1 and Cα2 isoforms. Cα3 isoform also arise as a result of alternative splicing of first coding exon with the internal exon. Newly identified transcript is expressed ubiquitously in different tissues examined. [ABSTRACT FROM AUTHOR]

The murine, bovine and human protein kinase A catalytic β ( Cβ) subunit genes encode several splice variants. Ten human Cβ gene splice variants, namely Cβ1, Cβ2, Cβ3, Cβ3b, Cβ3ab, Cβ3abc, Cβ4, Cβ4b, Cβ4ab and Cβ4abc, have been reported. Four splice variants of the murine Cβ gene are homologues of human Cβ1, Cβ2, Cβ3 and Cβ4 variants. Using a combinatorial approach comprising bioinformatics tools and molecular biology techniques, we have identified three novel alternatively spliced transcript variants of the mouse Cβ gene designated Cβ5, Cβ6 and Cβ7. These transcript variants differ in their first coding exon. New exons of Cβ5 and Cβ6 variants can encode different N-terminals; however, the new exon of variant Cβ7, when spliced with exon 2, encountered a stop codon in all three reading frames and thus cannot form a functional protein. The Cβ6 variant showed an ubiquitous pattern of expression, whereas Cβ5 was not expressed in muscle tissue on postnatal days (PN)3 and 15. Also, Cβ7 was found to be expressed only in brain and muscle tissues at PN3 and was absent in all tissues examined at PN15 and PN60. The post-translational modification analysis showed characteristic signature sequence motifs in predicted proteins important for the functionality of the protein. The human homologues of variants reported in the present study have not yet been identified. The present study reports three novel splice variants that have not been identified using conventional approaches of alternative splice variant detection methods. Therefore, the approach used in the present study can be used to identify splice variants of genes in organisms. Database [ABSTRACT FROM AUTHOR]