Abstract

This group has purified p57 and has obtained amino acid sequence from tryptic peptides. Degenerate oligonucleotides from these peptide sequences and from conserved sequences of serine/threonine kinases will be used to obtain PCR- generated DNA products.
In Aim 1, the PCR products will be ligated into pGEM3Zf-vector and the inserts sequenced to verify their difference from known MAP kinases. A unique sequence PCR product will be used to isolate a cDNA for p57 by screening a colon carcinoma cDNA library. Putative cDNA clones will be sequenced on both strands after insertion in both orientations in M13.
In Aim 2, they will determine whether the p57 MBP (myelin basic protein) kinase that is activated by TGFbeta1 in colon carcinoma cells is p57 MAP kinase. Polyclonal anti- peptide antisera to unique sequences (by GenBank analysis) in p57 MAP kinase will be generated and used to selectively remove p57 before analysis of TGFbeta1-modulated MAP kinases.
In Aim 3, they will determine whether Ki-ras, H-ras, PKCbeta, or c-src kinase play roles in p57 activation by transfection experiments. p57 MAP kinase is activated by DAGs in undifferentiated HT29 subclones that have elevated levels of both PKCbeta and c-src kinase. p57 MAP kinase cannot be activated by DAGs in differentiated HT29 subclones which exhibited about 10% of the PKCbeta activity or 20% of the c-src specific activity of undifferentiated subclones. Activities of c-src and PKC will be restored by transfection of expression plasmids for c-src, c-srcY527F, PKCbetaI, or PKCbetaII and p57 activation will be measured in the transfected cells.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA067405-03
Application #
2390878
Study Section
Special Emphasis Panel (ZRG3-ET-2 (01))
Project Start
1995-06-01
Project End
1999-03-31
Budget Start
1997-04-01
Budget End
1998-03-31
Support Year
3
Fiscal Year
1997
Total Cost
Indirect Cost

  Institution

Name
Upstate Medical University
Department
Pathology
Type
Schools of Medicine
DUNS #
058889106
City
Syracuse
State
NY
Country
United States
Zip Code
13210

  Publications

Ewton, Daina Z; Hu, Jing; Vilenchik, Maria et al. (2011) Inactivation of mirk/dyrk1b kinase targets quiescent pancreatic cancer cells. Mol Cancer Ther 10:2104-14
Hu, Jing; Friedman, Eileen (2010) Depleting Mirk Kinase Increases Cisplatin Toxicity in Ovarian Cancer Cells. Genes Cancer 1:803-811
Jin, Kideok; Ewton, Daina Z; Park, Sunju et al. (2009) Mirk regulates the exit of colon cancer cells from quiescence. J Biol Chem 284:22916-25
Deng, Xiaobing; Ewton, Daina Z; Friedman, Eileen (2009) Mirk/Dyrk1B maintains the viability of quiescent pancreatic cancer cells by reducing levels of reactive oxygen species. Cancer Res 69:3317-24
Jin, Kideok; Park, Sunju; Ewton, Daina Z et al. (2007) The survival kinase Mirk/Dyrk1B is a downstream effector of oncogenic K-ras in pancreatic cancer. Cancer Res 67:7247-55
Mercer, Stephen E; Ewton, Daina Z; Shah, Sejal et al. (2006) Mirk/Dyrk1b mediates cell survival in rhabdomyosarcomas. Cancer Res 66:5143-50
Deng, Xiaobing; Ewton, Daina Z; Li, Sheena et al. (2006) The kinase Mirk/Dyrk1B mediates cell survival in pancreatic ductal adenocarcinoma. Cancer Res 66:4149-58
Mercer, Stephen E; Friedman, Eileen (2006) Mirk/Dyrk1B: a multifunctional dual-specificity kinase involved in growth arrest, differentiation, and cell survival. Cell Biochem Biophys 45:303-15
Mercer, Stephen E; Ewton, Daina Z; Deng, Xiaobing et al. (2005) Mirk/Dyrk1B mediates survival during the differentiation of C2C12 myoblasts. J Biol Chem 280:25788-801
Deng, Xiaobing; Ewton, Daina Z; Mercer, Stephen E et al. (2005) Mirk/dyrk1B decreases the nuclear accumulation of class II histone deacetylases during skeletal muscle differentiation. J Biol Chem 280:4894-905

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