Sequence of a human brain adenylyl cyclase partial cDNA: Evidence for a consensus cyclase specific domain

doi:10.1016/0006-291X(91)91392-P

Biochemical and Biophysical Research Communications

Volume 179, Issue 1, 30 August 1991, Pages 455-462

https://doi.org/10.1016/0006-291X(91)91392-P Get rights and content

Abstract

A cDNA coding for a human brain adenylyl cyclase was isolated and sequenced. The deduced partial 675 amino-acid sequence was compared with those of other known adenylyl and guanylyl cyclases. Comparison of this predicted amino-acid sequence with that of bovine brain (type I) and rat olfactory (type III) adenylyl cyclase indicated a significant homology with the corboxyl-terminal halves of both enzymes. The homology between the human adenylyl cyclase and the other two mammalian adenylyl cyclase also appears at the topographic level. Indeed, the human enzyme includes an extremely hydrophobic region containing six potential membrane-spanning segments followed by a large hydrophilic domain. At the beginning of the hydrophilic domain, these is a 250 amino-acid region which shows not only a striking homology with the bovine and rat adenylyl cyclase (86% of similarity and 57% of identity), but also a significant homology with non-mammalian adenylyl cyclase and guanylyl cyclases. We found that this 250 amino-acid domain contains a sequence of about 165 amino-acid which is highly conserved in most of the known nucleotide cyclases suggesting that it includes residues that are critical for the function of the enzymes.

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Differential expression of adenylyl cyclase subtypes in human cardiovascular system
2004, Molecular and Cellular Endocrinology
In human myocardium, β₁-adrenoceptor stimulation achieves maximal inotropic response but less than 50% of maximal adenylyl cyclase activation, whereas the reverse is true of the β₂-adrenoceptor. Four types of adenylyl cyclase, type IV-VII, have been described in mammalian heart, but their expression and relative distribution in human heart and blood vessels is not known. We found that type IV, V, VI and VII adenylyl cyclases were all expressed in cardiomyocytes. Whereas types IV and VII RNA were more abundant in extra-cardiac than cardiac tissues, both absolute and relative expression of type VI was greatest in heart, and lower in tissues lacking a β₁-adrenoceptor. Type V expression was virtually confined to atrium. In situ mRNA hybridisation showed that the β₁-adrenoceptor co-localised with type VI adenylyl cyclase but not other subtypes in juxtoglomerular cells of human kidney. The tissue specific expression of these adenylyl cyclase subtypes may favour its coupling to corresponding receptors expressed in the given tissue type.
Quantitative reduction of type I adenylyl cyclase in human alcoholics
1999, Biochimica et Biophysica Acta - Molecular Basis of Disease
The amounts of adenylyl cyclase type I (AC I) were examined in various parts of the postmortem brains from alcoholics who prior to death had been abstinent from alcohol for at least 6 months and compared with controls using immunoblot analysis with anti-AC I specific antibody. It was revealed that a significant reduction of AC I was observed in both frontal and temporal cortices. On the other hand, in other areas (occipital cortex, caudate nucleus, putamen, and hippocampus) the amounts were comparable between alcoholics and controls. In the next step, we examined two subtypes of human AC mRNA levels (AC I and AC VIII) in blood cells by quantitative RT-PCR using [α-³²P]dCTP with two sets of the synthetic oligonucleotide primers based on the DNA sequences reported elsewhere (Villacres, E.C. et al., Genomics 16 (1993) 473–478; J. Parma et al., Biochem. Biophys. Res. Commun. 179 (1991) 455–462). The amounts of amplified DNAs of both AC I and AC VIII were significantly smaller in alcoholics than in controls. On the other hand, the amounts of amplified DNA of β-actin DNA were almost equal between alcoholics and controls. It appears from these results that a reduction in the amount of AC subtypes may be a biological marker for alcoholics.
Differential effect of chronic morphine on mRNA encoding adenylyl cyclase isoforms: Relevance to physiological sequela of tolerance/dependence
1998, Molecular Brain Research
In opiate naive longitudinal muscle myenteric plexus tissue, facilitation (G_S-mediated) and inhibition (G_i-mediated) of adenylyl cyclase (AC) activity is observed in response to low (nM) and high (μM) concentrations of sufentanil, respectively. Following chronic in vivo exposure to morphine, previously inhibitory concentrations produce excitatory effects. The present study was undertaken to explore the potential relevance of AC isoform-specific regulation to the qualitative change in opioid responsiveness following chronic morphine. Following persistent activation of opiate receptors, levels of AC I mRNA remain unchanged but that of AC IV is significantly augmented (approximately 37%; P<0.05). AC I and IV are differentially regulated by G_αi and G_βγ. The former is inhibited by G_αi and G_βγ whereas the latter is relatively insensitive to G_αi and is stimulated by G_βγ. Thus, an increase in AC IV mRNA could represent a shift from inhibitory to stimulatory opiate receptor-G protein signalling, as has been observed following chronic morphine. These results indicate that persistent activation of opiate receptors can induce selective changes in the abundance (activity) of AC isoforms. This could explain, in part, some of the adaptations that occur following chronic in vivo morphine exposure.
Splice variants of type VIII adenylyl cyclase: Differences in glycosylation and regulation by Ca<sup>2+</sup>/calmodulin
1996, Journal of Biological Chemistry
Three alternatively spliced type VIII adenylyl cyclase messages have been identified by cDNA cloning and amplification from rat brain cDNA. Type VIII-A was previously referred to simply as type VIII (Cali, J. J., Zwaagstra, J. C., Mons, N., Cooper, D. M. F., and Krupinski, J.(1994) J. Biol. Chem. 269, 12190-12195). The types VIII-B and -C cDNAs differ from that of type VIII-A by deletion of 90 and 198 base pair exons, respectively, which encode a 30-amino acid extracellular domain with two consensus sites for N-linked glycosylation and a 66-amino acid cytoplasmic domain. Stable expression of types VIII-A, -B, and -C cDNAs in human embryonal kidney 293 (HEK-293) cells leads to the appearance of novel proteins, which are recognized by type VIII-specific antibodies and which co-migrate with immunoreactive species detected on immunoblots of rat brain membranes. Types VIII-A and -C are modified by N-linked glycosylation, while type VIII-B is insensitive to treatment with N-glycosidase F. An influx of extracellular Ca²⁺ stimulates cAMP accumulation in HEK-293 cells stably expressing type VIII-A, -B, or -C, but not in control cells. Adenylyl cyclase activity of each of the variants is stimulated by Ca²⁺/calmodulin and the EC₅₀ for activation of type VIII-C is one fourth of that for either type VIII-A or -B. Type VIII-C also has a distinct K_m for substrate, which is approximately 4-12-fold higher than that for types VIII-A or -B depending on whether Mn²⁺ or Mg²⁺ is the counterion for ATP. The differences in the structural and enzymatic properties of these three variants are discussed.
[2] Analysis of adenylylcyclase subspecies gene expression in brain by in Situ hybridization histochemistry
1996, Methods in Neurosciences
This chapter presents the description of procedures that provide specific signals for the eight isoforms in areas of the rat brain and in individual neuronal cells and, thus, allows a fair estimation of mRNA levels for each adenylylcyclase isoform. In situ hybridization provides a useful tool for the identification of specific adenylylcyclase isoforms in the brain, and clearly demonstrates that the eight isoforms have a different pattern of expression. Fixation and sectioning are critical steps for the success of in situ hybridization. The restricted distributions of adenylylcyclase isoforms indicate that their expression is highly regulated for specific roles in the central nervous system. Because a number of the adenylylcyclases, closely related structurally, use the speciesspecific oligonucleotides and it appears to yield less ambiguous results, and more discrete signals are obtained with long riboprobes. It seems likely that this technique can be profitably applied to future studies on pharmacologically or developmentally induced changes in adenylylcyclase mRNA expression.
Identification and characterization of a widely expressed form of adenylyl cyclase
1996, Journal of Biological Chemistry
A novel mammalian adenylyl cyclase was identified by reverse transcription-polymerase chain reaction amplification using degenerate primers based on a conserved region of previously described adenylyl cyclases (Premont, R. T. (1994) Methods Enzymol. 238, 116-127). The full-length cDNA sequence obtained from mouse brain predicts a 1353-amino acid protein possessing a 12-membrane span topology, and containing two regions of high similarity with the catalytic domains of adenylyl cyclases. Comparison of this novel adenylyl cyclase with the eight previously described mammalian enzymes indicates that this type 9 adenylyl cyclase sequence is the most divergent, defining a sixth distinct subclass of mammalian adenylyl cyclases. The AC9 gene has been localized to human chromosome band 16p13.3-13.2. The 8.5-kb mRNA encoding the type 9 adenylyl cyclase is widely distributed, being readily detected in all tissues tested, and is found at very high levels in skeletal muscle and brain. AC9 mRNA is found throughout rat brain but is particularly abundant in hippocampus, cerebellum, and neocortex. An antiserum directed against the carboxyl terminus of the type 9 adenylyl cyclase detects native and expressed recombinant AC9 protein in tissue and cell membranes. Levels of the AC9 protein are highest in mouse brain membranes. Characterization of expressed recombinant AC9 reveals that the protein is a functional adenylyl cyclase that is stimulated by Mg²⁺, forskolin, and mutationally activated G_sα. AC9 activity is not affected by Ca²⁺/calmodulin or by G protein βγ-subunits. Thus AC9 represents a functional G protein-regulated adenylyl cyclase found in brain and in most somatic tissues.

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Sequence of a human brain adenylyl cyclase partial cDNA: Evidence for a consensus cyclase specific domain

Abstract

Trends Biochem. Sci

Methods Enzymol

Gene

J. Mol. Biol

Biochem. Biophys. Res. Commun

Febs Lett

J. Biol. Chem

Febs Lett

J. Biol. Chem

Cell

Cell

J. Biol. Chem

Cell

J. Biol. Chem

Gene

Febs Lett

J. Biol. Chem

Physiol. Rev

Annu. Rev. Biochem