Adenylyl cyclase activity of the fission yeast <i>Schizosaccharomyces pombe</i> is not regulated by guanyl nucleotides

Engelberg, David; Poradosu, Enrique; Simchen, Giora; Levitzki, Alexander

doi:10.1016/0014-5793(90)80605-i

Cited by 10 publications

(5 citation statements)

References 43 publications

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“…The evidence that GTP and its non-hydrolysable analogues affect cAMP production suggests that, in X marxianus, adenylate cyclase is regulated by GTP-binding proteins (Figure 2). In this respect the observed GTP-dependent activity points out a similarity with S. cerevisiae (Toda et al, 1985) rather than S. pombe (Engelberg et al, 1990a), as expected from systematics data (S. cerevisiae and Fl-D crude membranes (rasl:: URA3, ras2::LEUZ) were used as a negative control; we had similar results omitting proteins or antibodies from the immunoprecipitation assay. Sizes are indicated in kDa.…”

Section: Discussionsupporting

confidence: 80%

“…Mn2+dependent cAMP production was nearly eight-fold greater in K. marxianus than in S, cerevisiae SP1 strain (Gross et al, 1992; our unpublished results), but it was almost the same as in other S. cerevisiae strains (De Vendittis et al, 1986b;Becker et al, 1988). 30°C and pH 6.0 were assumed as standard conditions for all following experiments, considering that s. cerevisiae and s. pombe enzymes were characterized using the same conditions (Casperson et al, 1983;Engelberg et al, 1990a).…”

Section: K Marxianus Adenylate Cyclase Activitymentioning

confidence: 80%

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In the budding yeast Kluyveromyces marxianus, adenylate cyclase is regulated by Ras protein(s) in vitro

Verzotti

Geymonat

Valetti

et al. 1994

Yeast

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The presence of adenylate cyclase activity was first demonstrated in membrane fractions from the budding yeast Kluyveromyces marxianus. The enzyme showed a Mn(2+)- and Mg(2+)-dependent activity, with optimal pH at around 6 as observed in other yeast species. As in Saccharomyces cerevisiae, where adenylate cyclase is regulated by RAS1 and RAS2, we detected a guanyl nucleotide-dependent activity. Interestingly Y13-259 monoclonal antibody, raised against mammalian p21Ha-ras, inhibited Mg2+ plus GTP-gamma-S-dependent cAMP production, suggesting that the GTP binding proteins involved in adenylate cyclase regulation could be Ras proteins. The same antibody recognized on Western blot and immunoprecipitated a 40 kDa polypeptide from K. marxianus crude membranes. This polypeptide was not detected by an anti-RAS2 polyclonal antibody raised against S. cerevisiae RAS2 protein, suggesting that Ras proteins from the two species could be structurally different.

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Section: Discussionsupporting

confidence: 80%

Section: K Marxianus Adenylate Cyclase Activitymentioning

confidence: 80%

In the budding yeast Kluyveromyces marxianus, adenylate cyclase is regulated by Ras protein(s) in vitro

Verzotti

Geymonat

Valetti

et al. 1994

Yeast

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“…pombe is not activated by the addition of GTP in vitro, either in a membrane fraction (19,31) or in permeabilized cells (31), whereas this enzyme from S. cerevisiae can be activated (32). Engelberg et al (31) interpret this observation to suggest that Sch. pombe adenylyl cyclase is not regulated by G proteins.…”

Section: Resultsmentioning

confidence: 99%

Adenylyl cyclase is dispensable for vegetative cell growth in the fission yeast Schizosaccharomyces pombe.

Maeda

Mochizuki

Yamamoto

1990

Proc. Natl. Acad. Sci. U.S.A.

132

114

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Disruption of the cyri gene of Schizosaccharomyces pombe, which encodes adenylyl cyclase, did not confer lethality to fission yeast cells, although they grew 40% slower than wild-type strains in complete medium. These cells contained no measurable amount of cAMP and no adenylyl cyclase activity. When h' and h-cyri disruptants were mixed, they underwent mating even in rich medium. Propagation of homothallic cyri disruptants was difficult, probably because such cells readily mate and produce asci and thus stop growing. A >10-fold increase in the amount of cyri mRNA was observed when cloned cyrl+ was introduced into Sch. pombe cells on a multicopy plasmid. The total adenylyl cyclase activity was similarly high in these transformants. However, the level of intracellular cAMP was hardly affected. Evidence suggests that this was not due to increased phosphodiesterase activity. Thus, cAMP level in growing fission yeast cells appears to be regulated not by the amount of adenylyl cyclase protein but by a feedback mechanism at the enzyme level. The cAMP level fell by "50% under nitrogen starvation, which triggers sexual development in Sch. pombe. We suggest that fission yeast controls the level of intracellular cAMP primarily to regulate sexual development rather than to drive or arrest the cell cycle.

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“…The adenylate cyclase in vitro assays performed here almost certainly represent the unstimulated activity of adenylate cyclase and do not reflect the ability of adenylate cyclase to respond to in vivo stimuli. Adenylate cyclase activity from S. pombe is not stimulated by the addition of guanine nucleotides in vitro, and activity has only been observed when assayed in the presence of Mn2+; activity in the presence of Mg 2+ has not been detected (Yamawaki-Kataoka et al 1989;Engelberg et al 1990; C.S. Hoffman and F. Winston, unpubl.).…”

Section: Discussionmentioning

confidence: 99%

Glucose repression of transcription of the Schizosaccharomyces pombe fbp1 gene occurs by a cAMP signaling pathway.

Hoffman¹,

Winston²

1991

Genes Dev.

148

144

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Transcription of the fbpl gene, encoding fructose-l,6-bisphosphatase, of Schizosaccharomyces pombe is subject to glucose repression. Previous work has demonstrated that several genes {g/t genes} are required for this repression. In this report we demonstrate that one of these genes, git2, is the same as the cyrl gene, which encodes adenylate cyclase, and that loss-of-function mutations in git2 cause constitutive fbpl transcription. Addition of cAMP to the growth medium suppresses the transcriptional defect in git2 mutants as well as in strains that carry mutations in any of six additional g/t genes. Similarly, exogenous cAMP represses [bpl transcription in wild-type cells grown on a derepressing carbon source. Different levels of adenylate cyclase activity in different git2 mutants, coupled with the result that some git2 mutants display intragenic complementation, strongly suggest that adenylate cyclase acts as a multimer and that different git2 mutations alter distinct activities of adenylate cyclase, including catalytic activity and response to glucose.Additional experiments demonstrate that this cAMP signaling pathway is independent of the S. pombe rasl gene and works by activation of cAMP-dependent protein kinase.

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Adenylyl cyclase activity of the fission yeast Schizosaccharomyces pombe is not regulated by guanyl nucleotides

Cited by 10 publications

References 43 publications

In the budding yeast Kluyveromyces marxianus, adenylate cyclase is regulated by Ras protein(s) in vitro

In the budding yeast Kluyveromyces marxianus, adenylate cyclase is regulated by Ras protein(s) in vitro

Adenylyl cyclase is dispensable for vegetative cell growth in the fission yeast Schizosaccharomyces pombe.

Glucose repression of transcription of the Schizosaccharomyces pombe fbp1 gene occurs by a cAMP signaling pathway.

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