RESUMO
Dbl protein and DH (Dbl homology) domains are key regulators of RhoGTPases and promote GDP release from the complex with GTPase. About 70 DH-containing proteins are found. DH domain is localized in tandem with PH (pleckstrin homology) domain in many proteins. Bcr protein is a partner of Abl in reciprocal translocation t(9;22) which leads to Philadelphia chromosome formation. In the present study we have cloned Bcr DH and PH domains into the vector for mammalian expression. GEF activity of Bcr DH domain was studied alone and together with PH domain. Our data suggest, that Bcr DH domains does not reveal GEF activity against RhoGTPases RhoA, Cdc42 and Racl subfamilies in vivo.
Assuntos
Proteínas de Fusão bcr-abl/genética , Fatores de Troca do Nucleotídeo Guanina/metabolismo , Proteínas rho de Ligação ao GTP/metabolismo , Sequência de Aminoácidos , Linhagem Celular , Fatores de Troca do Nucleotídeo Guanina/genética , Humanos , Dados de Sequência Molecular , Cromossomo Filadélfia , Estrutura Terciária de Proteína , Homologia de Sequência de Aminoácidos , Transfecção , Proteínas rho de Ligação ao GTP/genéticaRESUMO
Ku protein is a multifunctional heterodimer of eukaryotes that consists of two subunits: Ku70 and Ku80. It plays a critical role in the regulation of many cellular processes such as: non-homologous end-joining, V(D)J-recombination of immunoglobulin genes and genes of T-cellular receptors, transcription regulation, DNA replication. Now the existence of relation between Ku protein functioning and processes that lead to transformation of normal cells into cancerous ones is determined, in Ph-positive leukemias and multiple myelomas, in particular. Nevertheless, the machinery of this processes is not still completely understood and needs further researches.
Assuntos
Antígenos Nucleares , Proteínas de Ligação a DNA , Neoplasias , Animais , Antígenos Nucleares/química , Antígenos Nucleares/genética , Antígenos Nucleares/fisiologia , Replicação do DNA/fisiologia , Proteínas de Ligação a DNA/química , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/fisiologia , Humanos , Autoantígeno Ku , Modelos Moleculares , Neoplasias/metabolismo , Neoplasias/patologia , Células Tumorais CultivadasRESUMO
Molecular-biological approaches was applied for detection of Philadelphia chromosome in patients with chronic myeloid and acute lymphoblastic leukemias. It includes bcr-genomic probes and polymerase chain reaction with specific bcr/abl primers. The latter is more preferable for clinic use. Different diagnostic methods are compared and leukemias etiology problems are discussed.
