Polyamine-activated protein kinase reaction from nuclei and nucleoli of Physarum polycephalum which phosphorylates a unique Mr 70,000 nonhistone protein

Daniels, Gary R.; Atmar, Valerie J.; Kuehn, Glenn D.

doi:10.1021/bi00512a025

Cited by 41 publications

(17 citation statements)

References 25 publications

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“…Polyamines have been reported to affect the activity of protein kinases from several organisms [17,IS]. In the case of the nuclear protein kinase from D. rliscoideum the influence of polyamines o n the enzyme activity has been found t o be quite complex.…”

Section: Substrate Specijkity Effect Of Camp and Polyaminesmentioning

confidence: 99%

Purification and properties of a cAMP‐independent nuclear protein kinase from Dictyostelium discoideum

Renart

Sastre

Sebastián

1984

European Journal of Biochemistry

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A cyclic-AMP-independent nuclear protein kinase has been purified from Dictyostelium discoideum amoebae. The purification procedure involves chromatography of DEAE-Sephadex, phosphocellulose and heparinSepharose. The purified enzyme phosphorylates threonine and serine of acidic proteins as casein and phosvitin. Phosphorylation of casein is stimulated by spermine. The kinase requires Mg2 + and can utilize both ATP and GTP as phosphoryl donors. Heparin is a potent inhibitor of the enzyme, being the protein kinase activity fully inhibited at concentrations of 0.5 pg/ml. One polypeptide of molecular mass 38 kDa was the major protein band present in the purified kinase preparation as estimated by NaDodSO, denaturing polyacrylamide gel electrophoresis. This band belongs to the protein kinase because it is the only one that is observed associated with the protein kinase activity when the enzyme preparation is centrifuged in glycerol gradients. The 38-kDa polypeptide is also the major product of autophosphorylation of the enzyme preparation. The enzymatic properties allow to classify the enzyme as a type-I1 casein kinase. However, its structural properties are different from the mammalian type-I1 casein kinases and make the D. discoideum enzyme more similar to the plants type-I1 casein kinases.Phosphorylation/dephosphorylation of proteins represents an important mechanism in the control of cellular processes, including the regulation of gene expression in eukaryotic cells I1,21.Among the enzymes that catalyze the postranslational phosphorylation of proteins are cyclic-nucleotide-dependent an independent protein kinases. Extensive investigations of CAMP-dependent protein kinases have shown that these enzymes mediate all of the biological effects of cyclic AMP. In contrast, little is known about the physiological role of the CAMP-independent protein kinases, although several types of these kind of enzyme have been characterized in different biological systems, mainly higher eukaryotes [3].The nuclear phosphorylation of chromatin-associated proteins, both histones and non histones, have been implicated in the regulation of gene transcription [4, 51. A cyclic-nucleotideindependent, polyamine-sensitive protein kinase has been involved in the control of transcription of ribosomal RNA genes by means of phosphorylation of RNA polymerase I [6] and some transcriptional factors [7].In the slime mold Dictyostelium discoideum a lot of effort has been dedicated to the study of the involvement of CAMPdependent protein kinases in the expression of its differentiation program. CAMP is the chemical agent that mediates aggregation of the amoebae [8] and it is also implicated in the regulation of latter stages of differentiation [9]. CAMPdependent protein kinases have been identified in D. discoideum both in vegetative cells [lo] and during differentiation [I 1, 121. Besides, the characterization of the regulatory subunit of a CAMP-dependent protein kinase has also been carried out using a photo-affinity probe [13]. However, the character...

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Section: Substrate Specijkity Effect Of Camp and Polyaminesmentioning

confidence: 99%

Purification and properties of a cAMP‐independent nuclear protein kinase from Dictyostelium discoideum

Renart

Sastre

Sebastián

1984

European Journal of Biochemistry

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“…Hence, the observed increase in 'IS incorporation into the 47 kD nuclear polypeptide could not be due to decreased phosphatase activity, but more likely is due to an increased kinase activity. Polyamines have been shown to regulate the activity of nuclear protein kinases in other systems (8,15,22).…”

Section: Discussionmentioning

confidence: 99%

Polyamine Stimulation of Protein Phosphorylation in Isolated Pea Nuclei

Datta

Hardison²,

Roux³

1986

Plant Physiol.

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ABSTRACIrThe phosphorylation of several proteins in isolated nuclei from Pisum sativum L. was stimulated by spermine. Although spermine increased the general protein phosphorylation by 10 to 20%, it increased the phosphorylation of a 47 kilodalton polypeptide by 150%. By comparison other polyamines, spermidine, putrescine, and cadavarine had far less effect on the phosphorylation of the 47 kilodalton or any other polypeptide. Sodium fluoride was able to inhibit the phosphorylation of the 47 kilodalton polypeptide in the control, implying the participation of protein phosphatase(s) in the phosphorylation of nuclear proteins. Spermine stimulated the phosphorylation of the 47 kilodalton polypeptide over the controls, even in the presence of NaF. This result indicates that spermine probably activates a nuclear kinase, a conclusion supported also by thiophosphorylation data. The inability of ethyleneglycol-bis(6-aminoethyl ether)-N,N'-tetraacetic acid and Compound 48/80, a calmodulin antagonist, to inhibit this spermine stimulated phosphorylation renders improbable any role ofcalcium and calmodulin in mediating this response.The role of polyamines in plant growth and development is well documented (11,25). Polyamines are known to regulate a wide variety of physiological and biochemical processes in the nuclei. Some of these include regulation of cell division (5, 16), and the activities of several enzymes involved in nucleic acid synthesis (14, 27) and repair (23). The activity of at least some of these enzymes is known to be controlled by polyamines via modulation of their phosphorylation state. In slime molds, Kuehn et al. (18) showed that polyamines regulate the phosphorylation of a certain nucleolar protein that regulates rRNA synthesis. They later identified this protein as ornithine decarboxylase and showed that polyamines act via regulating the activity of a protein kinase which in turn regulates the phosphorylation state and hence the activity of ornithine decarboxylase (2,19).We have already shown that phytochrome stimulates the phosphorylation of certain proteins in isolated nuclei from peas (9). In pea seedlings, phytochrome has also been shown to regulate polyamine biosynthesis by modulating the activity of arginine decarboxylase, one ofthe first enzymes in polyamine biosynthesis (6). In animal systems, polyamines are known to regulate nuclear protein phosphorylation (4,15,18). Given the above results, we decided to test the effect of polyamines on protein phosphorylation in isolated pea nuclei.In the present paper we show that polyamines, especially spermine, stimulate the phosphorylation of certain nuclear pro-'This work was supported by the National Science Foundation Grant PCM 8402526. teins in peas. We also present evidence that this stimulation is probably due to an increased activity of a protein kinase. Since the maximum stimulation of phosphorylation is of a 47 kD polypeptide most of the work in this paper is on the regulation of phosphorylation of the 47 kD polypeptide containing nuclear prote...

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“…It is well known that these hormones also stimulate the synthesis of rRNA at early times, but there have been conflicting reports about the role of the induced ODC in rRNA synthesis in nuclei. Kuehn and co-workers, who worked with Physarum polycephalum, reported that ODC, after being phosphorylated, is capable of stimulating the transcription of rRNA genes by RNA polymerase I (RNA nucleotidyltransferase, EC 2.7.7.6) (Kuehn et al, 1979;Daniels et al, 1981;. Russell and coworkers (Russell, 1981b;Russell & Manen, 1982) reported, using a rather crude preparation of ODC, that an ODC-putrescine conjugate could stimulate the activity of RNA polymerase I in liver nuclei and suggested that the conjugate is the S3 subunit of RNA polymerase I and the labile protein(s) itself which is supposed to mediate the hormonal regulation of rRNA.…”

Section: Introductionmentioning

confidence: 99%

Relationship between RNA polymerase I activity and ornithine decarboxylase in rat liver tissues

Urata¹,

Suzuki²,

Hosoya³

1987

Biochemical Journal

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In order to examine the relationship between RNA polymerase I and ornithine decarboxylase (ODC), three lines of experiments were performed, with the following results.(1) The glucocorticoid-induced increase of RNA polymerase I in rat liver nuclei was not abolished by administration of inhibitors of ODC synthesis and activity, namely 1,3-diaminopropane and 2-difluoromethylornithine respectively. (2) Anti-ODC antibody did not cross-react with RNA polymerase I solubilized from rat liver nucleoli, indicating the absence of a common protein sequence in these enzymes. (3) The ODC preparation which was treated with transglutaminase in the presence of putrescine could not stimulate the activity of RNA polymerase I in nuclei of liver and prostate. All these results suggest that the increases in ODC protein or activity are not a prerequisite to the increase in RNA polymerase I after hormonal or physiological stimuli, but rather that the increases in both enzymes are separate responses to the primary stimuli.

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Polyamine-activated protein kinase reaction from nuclei and nucleoli of Physarum polycephalum which phosphorylates a unique Mr 70,000 nonhistone protein

Cited by 41 publications

References 25 publications

Purification and properties of a cAMP‐independent nuclear protein kinase from Dictyostelium discoideum

Purification and properties of a cAMP‐independent nuclear protein kinase from Dictyostelium discoideum

Polyamine Stimulation of Protein Phosphorylation in Isolated Pea Nuclei

Relationship between RNA polymerase I activity and ornithine decarboxylase in rat liver tissues

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