A cytoplasmic cAMP-binding protein in Dictyostelium discoideum

Leichtling, Ben H.; Tihon, Claude; Spitz, Eva; Rickenberg, H.V.

doi:10.1016/0012-1606(81)90436-x

Cited by 21 publications

(7 citation statements)

References 25 publications

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“…Such sites seem to have been highly conserved through evolution since they also exhibit significant homology with the cAMP binding domain of the catabolite gene activator protein (CAP) from Escherichia coli (Weber et al, 1982). On the other hand, domains responsible for the interaction between regulatory and catalytic subunits of cAMP-dependent protein kinases seem to have also been highly conserved, since functional hybrid enzymes can be reconstructed by using catalytic subunit from bovine heart and regulatory subunits from bakers' yeast (Hixson & Krebs, 1980) or D. discoideum (Leichtling et al, 1981a;de Gunzburg & Veron, 1982). One could thus imagine that the regulatory subunit from D. discoideum that is monomeric and carries only one binding site for cAMP represents an "ancestral" form of the molecule.…”

Section: Discussionmentioning

confidence: 99%

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An unusual adenosine cyclic 3',5'-phosphate-dependent protein kinase from Dictyostelium discoideum

J¹,

Part²,

Guiso³

et al. 1984

Biochemistry

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The cAMP-dependent protein kinase from Dictyostelium discoideum was extracted from cells at the stage of culmination. Less than 50% of the enzyme remains as a cAMP-dependent holoenzyme in the extracts, and the rest is recovered in the form of dissociated regulatory and catalytic subunits that were purified. The regulatory subunit is a monomeric protein of Mr 42 000 that carries only one cAMP binding site (Xd = 3 nM). The catalytic subunit is also a monomer of M, 40000 with a sedimentation coefficient of 3.3 S. The cAMP-dependent holoenzyme is a dimer consisting

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Section: Discussionmentioning

confidence: 99%

“…Several cAMP binding proteins that could be intracellular targets for cAMP in D. discoideum cells have been described (Cooper et al, 1980; Leichtling et al, 1981a;de Gunzburg & Veron, 1981; Arents & van . Two types of proteins have been distinguished on the basis of their function, molecular structure, and developmental regulation.…”

mentioning

confidence: 99%

An unusual adenosine cyclic 3',5'-phosphate-dependent protein kinase from Dictyostelium discoideum

J¹,

Part²,

Guiso³

et al. 1984

Biochemistry

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“…The regulatory subunit of cAMP-dependent protein kinase is the best characterized cAMP binding protein in Dictyostelium (20)(21)(22)(23)(24)(25)(26)(27)(28)(29)(30). This protein has been suggested to play a role in gene regulation (30).…”

Section: Discussionmentioning

confidence: 99%

“…CABP1 is an unusual cAMP binding protein in that it consists oftwo subunits of different molecular weights. CABP2 has properties identical to the regulatory subunit of the cAMP-dependent protein kinase, which has been characterized extensively by other workers (20)(21)(22)(23)(24)(25)(26)(27)(28)(29)(30). The subcellular localization of the regulatory subunit of cAMP-dependent protein kinase has been examined before (22,30).…”

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confidence: 99%

Translocation of an unusual cAMP receptor to the nucleus during development of Dictyostelium discoideum.

Kay¹,

Noce²,

Tsang³

1987

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

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ABSTRACTcAMP has been implicated in the control of the expression of developmental genes in Dictyostelium discoi- We have recently characterized two cAMP binding proteins in developing D. discoideum cells, termed CABP1 and CABP2 (19). CABP1 is an unusual cAMP binding protein in that it consists oftwo subunits of different molecular weights. CABP2 has properties identical to the regulatory subunit of the cAMP-dependent protein kinase, which has been characterized extensively by other workers (20-30). The subcellular localization of the regulatory subunit of cAMP-dependent protein kinase has been examined before (22,30). In the present communication, we have examined the subcellular localization of CABP1 during development. CABP1 is found in the intracellular fraction as well as on the surface of developing cells. During development, CABP1 is translocated to the nucleus. This pattern of cellular localization is compatible with the idea that CABP1 plays a pivotal role in mediating the effect of cAMP on developmental gene expression. MATERIALS AND METHODSConditions of Growth and Development. The D. discoideum strain NC4 used in this study was originally isolated by K. Raper (Univ. of Wisconsin, Madison). The cells were grown in association with Enterobacter aerogenes on SM agar (31) in the dark at 220C for 40-44 hr until the bacterial lawn was just starting to clear. The vegetative amoebae were washed free of bacteria with 20 mM KH2PO4-K2HPO4 (pH 6.2) by centrifugation at 400 x g. The washed cells were used for analysis immediately if vegetative cells were required or allowed to develop at 5 x 106 cells per cm2 on 1.5% agar containing 40 mM KH2PO4-Na2HPO4 (pH 6.4), 20 mM KCl, and 2.5 mM MgCl2.Isolation of Nuclear and Postnuclear Supernatant Fractions. The isolation procedure was adapted from Cococci and Sussman (32). All steps were carried out at 40C unless otherwise specified. The cells were harvested at various times during development, washed twice with 0.2% NaCl, and resuspended at 108 cells per ml in NIB [50 mM HEPES, pH 7.5/5 mM magnesium acetate/10 mM benzamidine/50 ,ug of antipain per ml/50 pug of pepstatin per ml/2% Nonidet P-40/10% (wt/vol) sucrose]. An aliquot was removed and designated as total cellular extract.Abbreviation: CABP, cAMP binding protein.

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“…wt. protein that binds cAMP as well as adenosine and its derivatives is present in vegetative and aggregation competent cells (Gunzburg and Veron, 1981;Leichtling et al, 1981a). This protein, which is constitutive throughout the developmental cycle, will be described in *To whom reprint requests should be sent.…”

Section: Introductionmentioning

confidence: 99%

A cAMP-dependent protein kinase is present in differentiating Dictyostelium discoideum cells

Gunzburg

Véron

1982

The EMBO Journal

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We demonstrate the occurrence of a cAMP‐dependent protein kinase in Dictyostelium discoideum cells at the terminal stage of differentiation. A cAMP‐binding component was purified to homogeneity by affinity chromatography. This subunit inhibits the activity of purified catalytic subunit from beef heart protein kinase; the inhibition is reversed upon addition of cAMP. The protein is highly specific for cAMP and has a dissociation constant of 4 nM. The isolated regulatory subunit is a monomer of 39 K, with a sedimentation coefficient of 3.5S and a frictional coefficient of 1.24. The differences between this regulatory subunit and regulatory subunits of protein kinases from other sources are discussed.

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A cytoplasmic cAMP-binding protein in Dictyostelium discoideum

Cited by 21 publications

References 25 publications

An unusual adenosine cyclic 3',5'-phosphate-dependent protein kinase from Dictyostelium discoideum

An unusual adenosine cyclic 3',5'-phosphate-dependent protein kinase from Dictyostelium discoideum

Translocation of an unusual cAMP receptor to the nucleus during development of Dictyostelium discoideum.

A cAMP-dependent protein kinase is present in differentiating Dictyostelium discoideum cells

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