Cloning and sequence of partial cDNAs encoding the human type V and VI adenylyl cyclases and subsequent RNA-quantification in various tissues

Adenylyl cyclases (Acs) and VI are the predominant form of Acs in mammalian heart where they are part of the beta-adrenergic pathway. Up to now, the s equences for both enzymes from human tissues have not yet been reported. We investigated the mRNA expression AC V and VI in human colon, heart, liver, lung and MNL. Partial cDNAs of human types V and VI Acs were amplified by PCR using oligonucleotides derived from the cytoplasmatic domain sequences of the corresponding enzymes from dog heart. Primers derived from the human sequence were used to detect the mRNAs corresponding to both Acs. For quan tification of mRNAs we constructed internal standards for competitive quant itative reverse transcriptase PCR (RT-PCR). Both types of transcripts could be found in all investigated tissues except MNL where only type VI could b e detected. Further we demonstrated a more than 60 times higher amount of A C V-mRNA in human heart compared to AC VI-mRNA. (C) 1999 Elsevier Science B .V. All rights reserved.