The Human Gene for the Regulatory Subunit RI of Cyclic Adenosine 3',5'-Monophosphate-Dependent Protein Kinase: Two Distinct Promoters Provide Differential Regulation of Alternately Spliced Messenger Ribonucleic Acids

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Abstract
The present study reports the exon-intron organization of the hu- man RIa gene of cAMP-dependent protein kinase and approximately 2 kilobases (kb) of the 59-flanking region obtained by isolation and sequencing of several phage clones from human genomic libraries. The RIa gene is composed of nine coding exons of varying lengths, separated by introns, giving the gene a total length of at least 21 kb. Our recent cloning of a processed RIapseudogene with a 59-noncoding region different from the previously reported RIa complementary DNA indicated that the RIa gene may have multiple leader exons giving rise to alternately spliced messenger RNAs (mRNAs). Reverse transcription of human testis RNA followed by PCR identified two different RIa mRNA species (RIa1a and RIa1b) containing distinct 59-sequences due to alternately splicing the gene. The previously known RIa1b mRNA revealed low constitutive expression in a human B lymphoid cell line (Reh) and was stimulated only 4- to 6-fold by treatment with cAMP. In contrast, very low levels of the novel RIa1a mRNA were present in untreated Reh cells, but were stimulated 40- to 50-fold by cAMP. The 59-flanking sequence of the RIagene was G/C richanddidnotcontainanyTATAbox.Severalputativetranscription

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