Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving transferase
Patent
1997-06-19
1999-11-09
Carlson, Karen Cochrane
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving transferase
435194, C12Q 148, C12N 912
Patent
active
059812052
DESCRIPTION:
BRIEF SUMMARY
The current invention describes the identification of a novel widely-expressed human and D. melanogasterserine/threonine protein kinase (designated nuclear, Dbf2-related kinase, or Ndr; previously referred to as Pun kinase) and the use of this kinase for the identification of modulators thereof. The kinase is a nuclear protein and contains a short basic peptide, KRKAETWKRNRR, responsible for nuclear accumulation.
Reversible protein phosphorylation is a major mechanism for the coordinated control of many fundamental cellular functions in eukaryotic organisms, including metabolism, growth, and differentiation. The phosphorylation status, and consequently the activity, of specific target proteins is regulated by the opposing actions of protein kinases and protein phosphatases. Generally, these enzymes are specific either for serine/threonine or for tyrosine phosphoacceptors, although some dual specificity kinases and phosphatases have also been described. The importance of phosphorylation cascades is reflected by the finding that many kinases, phosphatases, and the signal transduction pathways in which they participate have been highly conserved during the course of evolution. In recent years, interest has focused on the role of protein phosphorylation in the control of the cell cycle; a number of cellular proto-oncogenes encode members of the serine/threonine kinase family and it has become increasingly clear that certain serine/threonine kinases function as key components of the cell cycle regulatory network. Therefore, the complete delineation of these pathways is an important aim for the understanding of oncogenesis and tumour progression.
The C. elegans expressed sequence tags (ESTs) cm11b7 and cm11b8 are overlapping cDNA clones which were originally described as worm homologues of the human kinase RAC/Akt (RAC-PK) and the S. cerevisiae cell cycle regulated kinase Dbf2 (Waterston et al., (1992) Nat. Genet. 1, 114-123). By complete sequencing of the clones, it has been determined that this homology assignment is incorrect.
Surprisingly it has been found that a novel kinase, distinct from RAC-PK and Dbf2, which is highly homologous to cm11b8 can be isolated from human and D. melanogaster sources. This kinase, Ndr, binds to calmodulin in a calcium-dependent manner. The gene encoding Ndr is located on human chromosome 6, between 6p21.2 and 6p21.31, a region of chromosome 6 which contains the major histocompatibility complex (MHC) class 1 genes. It has also been found that DNA encoding Ndr can be overexpressed when comprised in a suitable expression system, and that the protein sequence contains a short fragment that is responsible for the nuclear localisation of proteins.
SUMMARY OF THE INVENTION
In a first aspect of the present invention, we provide Ndr protein kinase, as well as homologues and derivatives thereof. Moreover, the invention relates to a nuclear localisation sequence derived from Ndr, and the use of Ndr for the modulation of calcium signalling.
DETAILED DESCRIPTION OF THE INVENTION
By conducting sequencing and homology studies using C. elegans cm11b7 and cm11b8 clones, we have determined that they represent a new protein kinase rather than a worm homologue of RAC-PK. The current invention concerns novel protein kinases comprising the amino acid sequence as given in SEQ ID NO:2 and SEQ ID NO:7 or a homologue thereof. The kinases of the invention are designated Ndr.
Ndr and its homologues are polypeptides which share sufficient similarities for the skilled person to determine that they share homology of origin or function with the Ndr protein kinases as represented by human and Drosophila Ndr. The invention includes all species homologues of Ndr. Human and Drosophila Ndr, represented in SEQ ID No. 7 and SEQ ID No. 2, are species homologues. Species homologues from other organisms may be isolated according to the methods set out herein, which are conventional. Moreover, suitable alternative methods are known to those of skill in the art and may be found in the literature. Species homologues
REFERENCES:
Cooper, J.A., et al., Meth. in Enzymol. 99:387-402 (1983).
Dang, C.V., et al., Mol. Cell Biol. 8(10):4048-4054 (1988). Abstract only.
Ellis, L., et al., Cell 45:721-732 (1986).
Hanks, S.K., et al., Meth. Enzymol. 200:38-62 (1991).
Hendrix, P., et al., J. Biol. Chem. 268(10):7330-7337 (1993).
Millward, T., et al., Experientia 50:A17 S05-24 (1994).
Millward, T., et al., Annual Report FMI 1993:44 (1995).
Millward, T., et al. PNAS USA 92:5022-5026 (1995).
Smith, D.B., et al., Gene. 67:31-40 (1988).
Waterston, R., et al., Nat. Genet. 1:114-123 (1992).
Millward et al., Poster presented at the USGEB meeting of Mar. 17th/18th 1994.
Dowell, S.J. et al. "Interaction of Dbf4, the Cdc7 protein kinase regulatory subunit, with yeast replication origins in vivo." Science (Aug. 1994), vol. 265, pp. 1243-1246.
Hemmings Brian Arthur
Millward Thomas Anders
Carlson Karen Cochrane
Ferraro Gregory D.
Novartis AG
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