Occurrence of NI and NII type protein kinases in the nuclei from various tissues of the rat

Yutani, Yasutaka; Tei, Yoko; Yukioka, Munehiko; Inoue, Akira

doi:10.1016/0003-9861(82)90362-9

Cited by 26 publications

(18 citation statements)

References 43 publications

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“…CK-2 has been reported to be present in the cytosol (23,64) and/or in the nucleus (31,65), depending on the cell type and on the experimental approaches used (discussed in reference 27). In rat liver cells, CK-2 is present in both the cytosol (28) and the nucleus (12,65).…”

Section: Calmodulin (Cam) Is a Ubiquitous Ca 2ϩmentioning

confidence: 99%

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Phosphorylation of Rat Liver Heterogeneous Nuclear Ribonucleoproteins A2 And C Can be Modulated by Calmodulin

Bosser

Faura

Serratosa

et al. 1995

Molecular and Cellular Biology

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). By immunoblotting, peptide mapping, and endogenous phosphorylation experiments, the 36-and 40-to 42-kDa proteins have been identified as the A2 and C proteins, respectively, of the heterogeneous nuclear ribonucleoprotein particles. To better understand the mechanism by which calmodulin inhibits the phosphorylation of these proteins, they were purified by using single-stranded DNA chromatography, and the effect of calmodulin on their phosphorylation by casein kinase 2 was analyzed. Results revealed that whereas calmodulin inhibited the phosphorylation of purified A2 and C proteins in a Ca 2؉ -dependent manner, it did not affect the casein kinase 2 phosphorylation of a different protein substrate, i.e., ␤-casein. These results indicate that the effect of calmodulin was not on casein kinase 2 activity but on specific protein substrates. The finding that the A2 and C proteins can bind to a calmodulin-Sepharose column in a Ca 2؉ -dependent manner suggests that this association could prevent the phosphorylation of the proteins by casein kinase 2. Immunoelectron microscopy studies have revealed that such interactions could also occur in vivo, since calmodulin and A2 and C proteins colocalize on the ribonucleoprotein particles in rat liver cell nuclei. Calmodulin (CaM) is a ubiquitous Ca 2ϩ-binding protein which has been highly conserved during evolution. The binding of Ca 2ϩ to CaM induces a conformational change which permits the interaction of CaM with specific target proteins, i.e., CaM-binding proteins. During the last few years, compelling evidence indicating that CaM is present in the cell nucleus (4) and that it has a role in several nuclear functions, such as gene transcription (21), DNA replication, DNA repair, and mitosis (see references 3, 4, and 36 for reviews), has accumulated.One of the ways to understand the molecular mechanisms by which CaM regulates these nuclear functions has been the identification of its nuclear protein targets. At least three groups of CaM-binding proteins have been found in the nuclei of different cell types: (i) proteins related to the actin motility systems, e.g., ␣-spectrin, myosin light-chain kinase, and caldesmon (6, 51, 57); (ii) transcription factors belonging to the basic helix-loop-helix structural group, e.g., human SEF-1, mouse E2A, and Max (21); and (iii) proteins involved in the phosphorylation or dephosphorylation of proteins, e.g., CaM-dependent protein kinase II and the CaM-dependent protein phosphatase calcineurin (11,51,54). The role of the specific complexes of CaM with these CaM-binding proteins in the nuclear functions still remains obscure, although some possible pathways have recently emerged. It has recently been reported that the addition of antiactin antibodies blocks the transport of macromolecules into purified nuclei in in vitro experiments, suggesting that the nuclear actin-myosin contractile system can play a role in nucleocytoplasmic transport (55). The transcription factors which are able to bind CaM and belong to the basic helix-loop-helix grou...

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Section: Calmodulin (Cam) Is a Ubiquitous Ca 2ϩmentioning

confidence: 99%

“…In rat liver cells, CK-2 is present in both the cytosol (28) and the nucleus (12,65). A broad range of CK-2 protein substrates has been identified.…”

Section: Calmodulin (Cam) Is a Ubiquitous Ca 2ϩmentioning

confidence: 99%

Phosphorylation of Rat Liver Heterogeneous Nuclear Ribonucleoproteins A2 And C Can be Modulated by Calmodulin

Bosser

Faura

Serratosa

et al. 1995

Molecular and Cellular Biology

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“…5 b). Furthermore, when S1 proteins were incubated with [p3'P]ATP together with highly purified protein kinase NII (a major protein kinase in the cell nuclei of most tissues [16]), the same spots as well as three additional spots of D2 proteins and three spots of C2 proteins were found to be phosphorylated by the enzyme (Fig. 5 b).…”

Section: (-)mentioning

confidence: 99%

“…The enzyme activities of protein kinase, poly(ADPribose) polymerase and RNA polymerase I1 were assayed as previously described in [16], [17] and [I 81, respectively.…”

Section: Assay For Enzyme Activitiesmentioning

confidence: 99%

A set of non‐histone proteins isolated from the nuclei of various rat tissues

Inoue

Higashi

Hasuma

et al. 1983

European Journal of Biochemistry

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A set of non-histone proteins has been identified in the nuclei from liver, brain, spleen and testis tissues of the rat. Following moderate digestion of thoroughly washed nuclei with DNase 1 or micrococcal nuclease, EDTA was added to 5 m M to the reaction mixture and the preparation centrifuged. We found that the supernatant contained a limited amount of non-histone proteins (fraction SI). Sodium dodecyl sulfate (SDS) gel electrophoresis revealed S1 to be composed of a remarkably simple set of proteins resolved into four groups (A -D) each possessing closely spaced doublets or a triplet. Their molecular weights were A, 76100-80000; B, 48200-49500; C, 44500-45200 and D, 39500-41 500. The yield suggested that these proteins were structural constituents; however, they did not coincide with the known structural proteins of the cell nucleus. Two-dimensional gel electrophoresis further resolved each of the SDS bands into as many as nine spots, according to various charges. Some were labelled with [32P]orthophosphate in vivo, or with [y-32P]ATP and purified nuclear protein kinase NII in vitro. The released proteins B -D had fairly constant relative molar ratios at various times of digestion, thereby indicating possible localizations a t similar sites in the nucleus. The kinetic data together with the aggregation property at neutral pH values and the solubility in 5 mM EDTA suggest that proteins B -D constitute a group of proteins that have several common characteristics.The abundance of protein species in the cell nucleus varies with the protein; some are structural or regulatory in function while others are catalytic in various biochemical processes. Isolation and characterization of these nuclear proteins should greatly aid in understanding the structure and function of the nucleus as well as the molecular properties of the protein in question. Isolation of a group of proteins which share molecular properties may be advantageous, as a common biological function may be involved. The precedent of this can be illustrated with histones and high mobility group (HMG) proteins. Histones, the building molecules of the nucleosomes [I], can be readily extracted in a set from the chromatin with dilute mineral acids [2]; H M G proteins, which confer a structural characteristic of the active Chromatin [3], are extracted as a group from chromatin with 0.35 M NaCl and remain soluble in 2There have been numerous studies on various nuclear architectures such as chromatin, nuclear matrix and nuclear envelopes. One approach is to use DNA-hydrolyzing enzymes ; for example, DNase I has been used to distinguish the transcriptionally active from the bulk chromatin in differential digestion kinetics [5], or to isolate the nuclear pore complexlamina fraction [6] and nuclear matrix [7].In our studies on nuclear proteins we were interested in protein components liberated from the nuclei by digestion with DNA-hydrolyzing enzymes. We report here that a novel and remarkably simple set of non-histone proteins was isolated from the nuclei by nucle...

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“…Since some protein kinases are affected by polyamines (Glover et al, 1983;Tabor & Tabor, 1976) and heparin (Hathaway et af., 1980;Yutani et al, 1982), the effects of these compounds were examined using mixed histones as the phosphate acceptor. PK I1 activity was stimulated about l.5-fold by 0-5 mM-spermine or spermidine, and threefold when the concentration of either of these polyamines in the assay medium was 2 mM.…”

Section: A K S a H A A N D O T H E R Smentioning

confidence: 99%

Demonstration of Two Protein Kinases in Extracts of Legionella micdadei

Saha

Dowling²,

Mukhopadhyay³

et al. 1988

Microbiology

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Protein kinases I (PK I) and I1 (PK 11) were purified 253-and 13.5-fold, respectively, from an extract of sonically disrupted cells of Legionella micdadei by ion-exchange chromatography on QAE-Sephadex, by histone affinity chromatography, and by HPLC-gel filtration chromatography. Both enzymes catalysed the phosphorylation of calf thymus histones, with a K , of 2.7mgml-I for PK I and 2.9 mg ml-l for PK 11. Histone HZb was the best protein kinase substrate for both PK I and PK 11. The pH optima were 6.8 and 7.0 for PK I and PK I1 respectively. The K , for ATP was 0-29 mM for PK I and 0.33 mM for PK 11. PK I1 activity was stimulated by either CAMP or cGMP, whereas PK I was inhibited by both cyclic nucleotides. The activity of PK I was unaffected by addition of calmodulin, diacylglycerol and mixtures of Ca2+ and acidic phospholipids, but these additions increased PK I1 activity threefold. The activity of PK I1 was stimulated by spermine and spermidine, but PK I was inhibited by these compounds. PK I and PK I1 were both strongly inhibited by heparin.

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Occurrence of NI and NII type protein kinases in the nuclei from various tissues of the rat

Cited by 26 publications

References 43 publications

Phosphorylation of Rat Liver Heterogeneous Nuclear Ribonucleoproteins A2 And C Can be Modulated by Calmodulin

Phosphorylation of Rat Liver Heterogeneous Nuclear Ribonucleoproteins A2 And C Can be Modulated by Calmodulin

A set of non‐histone proteins isolated from the nuclei of various rat tissues

Demonstration of Two Protein Kinases in Extracts of Legionella micdadei

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