Cloning of a cdc2-related protein kinase from Trypanosoma cruzi that interacts with mammalian cyclins

Mol Biochem Parasitol. 1998 Mar 15;91(2):337-51. doi: 10.1016/s0166-6851(97)00218-1.

Abstract

Two cdc2-related protein kinases (crk), tzcrk3 and tzcrk1, from the protozoan parasite Trypanosoma cruzi were cloned. tzcrk3 encodes a 35 kDa protein sharing 51.5% amino acid identity with human cdc2 and 82% identity with Trypanosoma brucei CRK3. tzcrk1 encodes a 33 kDa protein sharing 52.7% identity with human cdc2 and a high degree of identity (> 78%) with T. brucei CRK1, Leishmania mexicana CRK1 and Trypanosoma congolense CRK1. A recombinant TzCRK1 protein was able to phosphorylate histone HI and retinoblastoma protein. Western blotting using a polyclonal antibody raised against the recombinant TzCRK1 protein showed that the kinase is present in all life cycle stages of the parasite. A PSTAIRE antiserum detected proteins of 32, 33 and 35 kDa, with differential expression in the life cycle of the parasite. Transfection of COS-7 cells with tzcrk1 demonstrated for the first time that a CRK protein can bind mammalian cyclins; TzCRK1 co-immunoprecipitated with cyclins E, D3 and A suggesting a role for this kinase in cell cycle control. These results indicate that T. cruzi might have cyclin homologues that control the activity of the CRK proteins and that a complex mechanism would exist in order to regulate the kinases involved in the cell cycle and the differentiation processes of the parasite.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Blotting, Southern
  • CDC2 Protein Kinase / chemistry
  • CDC2 Protein Kinase / genetics
  • CDC2 Protein Kinase / metabolism
  • COS Cells
  • Cloning, Molecular
  • Cyclins / metabolism*
  • Genes, Protozoan
  • Genetic Complementation Test
  • Histones / metabolism
  • Humans
  • Molecular Probe Techniques
  • Molecular Sequence Data
  • Protein Kinases / chemistry
  • Protein Kinases / genetics*
  • Protein Kinases / metabolism
  • Protozoan Proteins / chemistry
  • Protozoan Proteins / genetics*
  • Protozoan Proteins / metabolism
  • Recombinant Proteins / metabolism
  • Retinoblastoma Protein / metabolism
  • Sequence Alignment
  • Transfection
  • Trypanosoma cruzi / enzymology*
  • Trypanosoma cruzi / genetics
  • Trypanosoma cruzi / growth & development

Substances

  • CRK protein, protozoan
  • Cyclins
  • Histones
  • Protozoan Proteins
  • Recombinant Proteins
  • Retinoblastoma Protein
  • Protein Kinases
  • TCRK3 protein, Trypanosoma cruzi
  • tcrk1 protein, Trypanosoma cruzi
  • CDC2 Protein Kinase

Associated data

  • GENBANK/U69958
  • GENBANK/U69959
  • GENBANK/U69960
  • GENBANK/U74762
  • GENBANK/U74763
  • GENBANK/U74764
  • GENBANK/U74765
  • GENBANK/U74766