cDNA cloning and characterization of rat Clk3, a LAMMER kinase predominately expressed in testis

Becker, Walter; Kentrup, H.; Heukelbach, Jörg; Joost, Hans‐Georg

doi:10.1016/0167-4889(96)00036-5

Cited by 20 publications

(17 citation statements)

References 12 publications

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“…With all kinases of the DYRK family, incorporation of 32 P into H3 was greatly reduced or completely abolished in the mutant protein, clearly indicating that Thr-45 is the main phosphorylation site in histone H3 for DYRK-related kinases. For comparison, GST-CLK3 was included as a more distantly related protein kinase (18). Large amounts of the kinase were required to achieve detectable phosphorylation of rH3, and no difference was observed between rH3 and rH3-T45A (Fig.…”

Section: Resultsmentioning

confidence: 99%

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Specificity Determinants of Substrate Recognition by the Protein Kinase DYRK1A

Himpel¹,

Tegge²,

Frank³

et al. 2000

Journal of Biological Chemistry

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223

221

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DYRK1A is a dual-specificity protein kinase that is thought to be involved in brain development. We identified a single phosphorylated amino acid residue in the DYRK substrate histone H3 (threonine 45) by mass spectrometry, phosphoamino acid analysis, and protein sequencing. Exchange of threonine 45 for alanine abolished phosphorylation of histone H3 by DYRK1A and by the related kinases DYRK1B, DYRK2, and DYRK3 but not by CLK3. In order to define the consensus sequence for the substrate specificity of DYRK1A, a library of 300 peptides was designed in variation of the H3 phosphorylation site. Evaluation of the phosphate incorporation into these peptides identified DYRK1A as a proline-directed kinase with a phosphorylation consensus sequence (RPX(S/T)P) similar to that of ERK2 (PX(S/T)P). A peptide designed after the optimal substrate sequence (DYRKtide) was efficiently phosphorylated by DYRK1A (K m ‫؍‬ 35 M) but not by ERK2. Both ERK2 and DYRK1A phosphorylated myelin basic protein, whereas only ERK2, but not DYRK1A, phosphorylated the mitogenactivated protein kinase substrate ELK-1. This marked difference in substrate specificity between DYRK1A and ERK2 can be explained by the requirement for an arginine at the P ؊3 site of DYRK substrates and its presumed interaction with aspartate 247 conserved in all DYRKs.DYRK1A is a nuclear protein kinase that is ubiquitously expressed in rat tissues (1, 2). We have recently characterized a closely related isoform, DYRK1B, that is predominantly expressed in testis and muscle (3). The homolog of DYRK1A and DYRK1B in Drosophila, the protein kinase MNB, is encoded by the minibrain gene whose mutation results in specific defects in the development of the central nervous system (4). The human gene for DYRK1A is located in the "Down syndrome critical region" of chromosome 21, and the similarity of DYRK1A and MNB suggests that the triplication of the DYRK1A gene may play a role in mental retardation of patients with Down syndrome (5-9).DYRK1A, DYRK1B, and MNB belong to a subfamily of protein kinases with structurally related catalytic domains and similar enzymatic properties (2, 10). At least 7 different DYRKrelated kinases exist in mammals, of which DYRK1A and DYRK1B are targeted to the nucleus, whereas DYRK2 and DYRK3 are located in the cytoplasm (2, 3).1 Members of the DYRK family have also been found in lower eukaryotes, such as Yak1p in Saccharomyces cerevisiae (11), Pom1p in Schizosaccharomyces pombe (12), and YAKA in Dictyostelium discoideum (13). Although mutations in YAK1, pom1, and yakA have diverse phenotypic consequences, it appears reasonable to generalize that DYRK-related kinases are involved in the regulation of growth and development.The enzymatic activity of DYRK1A has been shown to depend on the presence of tyrosine residues in the activation loop, a result that suggests an activation mechanism similar to that of the MAP 2 kinases (1). However, the participation of DYRK1A or other DYRK-related kinases in a particular signal transduction pathway has not been eluci...

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Section: Resultsmentioning

confidence: 99%

“…Protein Kinases and Substrates-The expression plasmids for GST-DYRK1A, GST-DYRK2, GST-DYRK3, and GST-CLK3 have been described earlier (1,2,18). An NcoI site at base pair 1624 of the rat cDNA was used to create a C-terminal deletion (amino acids 500 -763, numbering refers to Swiss-Prot accession number Q63470) in GST-DYRK1A-⌬C.…”

Section: Methodsmentioning

confidence: 99%

Specificity Determinants of Substrate Recognition by the Protein Kinase DYRK1A

Himpel¹,

Tegge²,

Frank³

et al. 2000

Journal of Biological Chemistry

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223

221

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“…mRNA expression revealed by Northern blot analysis showed that PRPK may be functioning considerably in the testes in vivo. Testis-specific protein kinases have been reported (22)(23)(24). Both tssk1 and tssk2 appear in the adult testis and may play a role in the last stage of spermatid maturation (24).…”

Section: Discussionmentioning

confidence: 99%

Cloning and Characterization of a p53-related Protein Kinase Expressed in Interleukin-2-activated Cytotoxic T-cells, Epithelial Tumor Cell Lines, and the Testes

Abe¹,

Matsumoto²,

Wei³

et al. 2001

Journal of Biological Chemistry

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A human protein kinase, p53-related protein kinase (PRPK), was cloned from an interleukin-2-activated cytotoxic T-cell subtraction library. PRPK appears to be a homologue of a growth-related yeast serine/threonine protein kinase, YGR262c. However, a complementation assay using YGR262c-disrupted yeast indicated that PRPK is not functionally identical to the yeast enzyme. PRPK expression was observed in interleukin-2-activated cytotoxic T-cells, some human epithelial tumor cell lines, and the testes. The intrinsic transcriptional activity of p53 was up-regulated by a transient transfection of PRPK to COS-7 cells. PRPK was shown to bind to p53 and to phosphorylate p53 at Ser-15. These results indicate that PRPK may play an important role in the cell cycle and cell apoptosis through phosphorylation of p53.

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“…In normal tissues, only the testis expresses TOPK mRNA. Testis-specific protein kinases have been reported (27)(28)(29)(30), and some spermatogenetic roles of these kinases have been debated; however, the precise mechanisms of these kinases, such as target specificity and signaling cascades, remain to be elucidated. On the other hand, TOPK phosphorylates p38 MAPK, and its signaling can be examined from the viewpoint of a MAPK cascade in the testis.…”

Section: Fig 12mentioning

confidence: 99%