Identification of the domain of Saccharomyces cerevisiae adenylate cyclase associated with the regulatory function of RAS products

1987 ◽  
Vol 210 (2) ◽  
pp. 187-194 ◽  
Author(s):  
Isao Uno ◽  
Hiroshi Mitsuzawa ◽  
Kazuma Tanaka ◽  
Takehiro Oshima ◽  
Tatsuo Ishikawa
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Adenylate Cyclase ◽  
Regulatory Function

Regulatory function of the Saccharomyces cerevisiae RAS C-terminus

1987 ◽  
Vol 7 (7) ◽  
pp. 2309-2315
Author(s):  
M S Marshall ◽  
J B Gibbs ◽  
E M Scolnick ◽  
I S Sigal
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Adenylate Cyclase ◽  
Attachment Site ◽  
Hydrolytic Activity ◽  
Regulatory Function ◽  
Coding Region ◽  
Activating Mutations ◽  
Terminal Domain ◽  
C Terminus ◽  
Type Gene

Activating mutations (valine 19 or leucine 68) were introduced into the Saccharomyces cerevisiae RAS1 and RAS2 genes. In addition, a deletion was introduced into the wild-type gene and into an activated RAS2 gene, removing the segment of the coding region for the unique C-terminal domain that lies between the N-terminal 174 residues and the penultimate 8-residue membrane attachment site. At low levels of expression, a dominant activated phenotype, characterized by low glycogen levels and poor sporulation efficiency, was observed for both full-length RAS1 and RAS2 variants having impaired GTP hydrolytic activity. Lethal CDC25 mutations were bypassed by the expression of mutant RAS1 or RAS2 proteins with activating amino acid substitutions, by expression of RAS2 proteins lacking the C-terminal domain, or by normal and oncogenic mammalian Harvey ras proteins. Biochemical measurements of adenylate cyclase in membrane preparations showed that the expression of RAS2 proteins lacking the C-terminal domain can restore adenylate cyclase activity to cdc25 membranes.

scholarly journals Regulatory function of the Saccharomyces cerevisiae RAS C-terminus.

1987 ◽  
Vol 7 (7) ◽  
pp. 2309-2315 ◽  
Author(s):  
M S Marshall ◽  
J B Gibbs ◽  
E M Scolnick ◽  
I S Sigal
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Adenylate Cyclase ◽  
Attachment Site ◽  
Hydrolytic Activity ◽  
Regulatory Function ◽  
Coding Region ◽  
Activating Mutations ◽  
Terminal Domain ◽  
C Terminus ◽  
Type Gene

Activating mutations (valine 19 or leucine 68) were introduced into the Saccharomyces cerevisiae RAS1 and RAS2 genes. In addition, a deletion was introduced into the wild-type gene and into an activated RAS2 gene, removing the segment of the coding region for the unique C-terminal domain that lies between the N-terminal 174 residues and the penultimate 8-residue membrane attachment site. At low levels of expression, a dominant activated phenotype, characterized by low glycogen levels and poor sporulation efficiency, was observed for both full-length RAS1 and RAS2 variants having impaired GTP hydrolytic activity. Lethal CDC25 mutations were bypassed by the expression of mutant RAS1 or RAS2 proteins with activating amino acid substitutions, by expression of RAS2 proteins lacking the C-terminal domain, or by normal and oncogenic mammalian Harvey ras proteins. Biochemical measurements of adenylate cyclase in membrane preparations showed that the expression of RAS2 proteins lacking the C-terminal domain can restore adenylate cyclase activity to cdc25 membranes.

The activation of adenylate cyclase by guanyl nucleotides in Saccharomyces cerevisiae is controlled by the CDC25 start gene product

1987 ◽  
Vol 7 (10) ◽  
pp. 3857-3861
Author(s):  
J Daniel ◽  
J M Becker ◽  
E Enari ◽  
A Levitzki
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Adenylate Cyclase ◽  
Gene Product ◽  
Ternary Complex ◽  
Restrictive Temperature ◽  
Permissive Temperature ◽  
Terminal Portion ◽  
Altered Regulation ◽  
Cdc25 Protein

In the thermosensitive cdc25 start mutant of Saccharomyces cerevisiae, the regulation of adenylate cyclase by guanyl nucleotides was rapidly nullified when the enzyme was prepared from nonsynchronized cells shifted to the restrictive temperature. In agreement with previous in vivo complementation studies, this biochemical defect was fully suppressed by the expression of either the whole cloned CDC25 gene or its C-terminal portion. Moreover, membranes prepared from cdc25(Ts) cells grown at the permissive temperature evinced an altered regulation of adenylate cyclase by guanyl nucleotides. These results indicate that the CDC25 protein, together with RAS, is involved in the regulation of adenylate cyclase by guanyl nucleotides and raise the possibility that adenylate cyclase might form a ternary complex with RAS and CDC25.

Mutational analysis of the Saccharomyces cerevisiae SNF1 protein kinase and evidence for functional interaction with the SNF4 protein

1989 ◽  
Vol 9 (11) ◽  
pp. 5034-5044
Author(s):  
J L Celenza ◽  
M Carlson
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Protein Kinase ◽  
Mutational Analysis ◽  
Gene Dosage ◽  
Regulatory System ◽  
Kinase Domain ◽  
Regulatory Function ◽  
Protein Kinase Activity ◽  
Glucose Limitation

The SNF1 gene of Saccharomyces cerevisiae encodes a protein-serine/threonine kinase that is required for derepression of gene expression in response to glucose limitation. We present evidence that the protein kinase activity is essential for SNF1 function: substitution of Arg for Lys in the putative ATP-binding site results in a mutant phenotype. A polyhistidine tract near the N terminus was found to be dispensable. Deletion of the large region C terminal to the kinase domain only partially impaired SNF1 function, causing expression of invertase to be somewhat reduced but still glucose repressible. The function of the SNF4 gene, another component of the regulatory system, was required for maximal in vitro activity of the SNF1 protein kinase. Increased SNF1 gene dosage partially alleviated the requirement for SNF4. C-terminal deletions of SNF1 also reduced dependence on SNF4. Our findings suggest that SNF4 acts as a positive effector of the kinase but does not serve a regulatory function in signaling glucose availability.

RPD3 encodes a second factor required to achieve maximum positive and negative transcriptional states in Saccharomyces cerevisiae

1991 ◽  
Vol 11 (12) ◽  
pp. 6317-6327 ◽  
Author(s):  
M Vidal ◽  
R F Gaber
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Target Genes ◽  
Current Data ◽  
Regulatory Function ◽  
Fourfold Increase ◽  
Recessive Mutations ◽  
Activation Of Transcription ◽  
Transcriptional Regulatory ◽  
Low Levels ◽  
Full Activation

In Saccharomyces cerevisiae, TRK1 and TRK2 encode the high- and low-affinity K+ transporters, respectively. In cells containing a deletion of TRK1, transcription levels of TRK2 are extremely low and are limiting for growth in media containing low levels of K+ (Trk- phenotype). Recessive mutations in RPD1 and RPD3 suppress the TRK2, conferring an approximately fourfold increase in transcription. rpd3 mutations confer pleiotropic phenotypes, including (i) mating defects, (ii) hypersensitivity to cycloheximide, (iii) inability to sporulate as homozygous diploids, and (iv) constitutive derepression of acid phosphatase. RPD3 was cloned and is predicted to encode a 48-kDa protein with no extensive similarity to proteins contained in current data bases. Deletion of RPD3 is not lethal but confers phenotypes identical to those caused by spontaneous mutations. RPD3 is required for both full repression and full activation of transcription of target genes including PHO5, STE6, and TY2. RPD3 is the second gene required for this function, since RPD1 is also required. The effects of mutations in RPD1 and RPD3 are not additive, suggesting that these genes are involved in the same transcriptional regulatory function or pathway.

Properties and Possible Functions of the Adenylate Cyclase in Plasma Membranes of Saccharomyces cerevisiae

1982 ◽  
Vol 2 (12) ◽  
pp. 1481-1491
Author(s):  
Patrick K. Jaynes ◽  
James P. McDonough ◽  
Henry R. Mahler
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Protein Kinase ◽  
Adenylate Cyclase ◽  
Plasma Membranes ◽  
Dissociation Constants ◽  
Guanine Nucleotides ◽  
Adenylate Cyclase System ◽  
Regulatory Subunits ◽  
Molecular Weights ◽  
Essential Components

We have examined the possible role of adenosine 3′,5′-phosphate (cAMP) in functions associated with the plasma membranes of Saccharomyces cerevisiae. Purified membranes from this source contained an adenylate cyclase which was insensitive to activation by fluoride or guanine nucleotides, only weakly responsive to changes of carbon source in the growth medium, and strongly stimulated by vanadate. They also contained at least two classes of receptor proteins for guanine nucleotides (as measured by binding of labeled 5′-guanylyl methylene diphosphate) with apparent dissociation constants equal to 1.0 × 10 −7 and 3 × 10 −6 M, a protein kinase capable of phosphorylating added histones, the activity of which was stimulated by cAMP, and cAMP receptors that may function as regulatory subunits for this kinase. Membrane proteins were also susceptible to phosphorylation by endogenous kinase(s), with polypeptides of apparent molecular weights equal to 160 × 10 3 , 135 × 10 3 , 114 × 10 3 , and 58 × 10 3 as the major targets. Of these, the 114,000-molecular-weight polypeptide was probably identical to the proton-translocating ATPase of the membranes. However, the cAMP-dependent protein kinase did not appear to be involved in these reactions. Intact ( rho + or rho 0 ) cells responded to dissipation of the proton electrochemical gradient across their plasma membranes by rapid and transient changes in their intracellular level of cAMP, as suggested earlier (J. M. Trevillyan and M. L. Pall, J. Bacteriol., 138 :397-403, 1979). Thus, although yeast plasma membranes contain all the essential components of a stimulus-responsive adenylate cyclase system, the precise nature of the coupling device and the targets involved remain to be established.

scholarly journals Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP) in Physiological and Pathological Processes within the Gastrointestinal Tract: A Review

2021 ◽  
Vol 22 (16) ◽  
pp. 8682
Author(s):  
Aleksandra Karpiesiuk ◽  
Katarzyna Palus
Keyword(s):  
Adenylate Cyclase ◽  
Smooth Muscle Contraction ◽  
Regulatory Function ◽  
Gi Tract ◽  
Cytoprotective Effect ◽  
Neoplastic Processes ◽  
Pituitary Adenylate Cyclase ◽  
Pleiotropic Action ◽  
Tract Function ◽  
The Central Nervous System

Pituitary adenylate cyclase-activating polypeptide (PACAP) is a neuropeptide widely distributed in the central nervous system (CNS) and many peripheral organs, such as the digestive tract, endocrine, reproductive and respiratory systems, where it plays different regulatory functions and exerts a cytoprotective effect. The multifarious physiological effects of PACAP are mediated through binding to different G protein-coupled receptors, including PAC1 (PAC1-R), VPAC1 (VPAC1-R) and VPAC2 (VPAC2-R) receptors. In the gastrointestinal (GI) tract, PACAP plays an important regulatory function. PACAP stimulates the secretion of digestive juices and hormone release, regulates smooth muscle contraction, local blood flow, cell migration and proliferation. Additionally, there are many reports confirming the involvement of PACAP in pathological processes within the GI tract, including inflammatory states, neuronal injury, diabetes, intoxication and neoplastic processes. The purpose of this review is to summarize the distribution and pleiotropic action of PACAP in the control of GI tract function and its cytoprotective effect in the course of GI tract disorders.

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