RESUMO
Background: TRF2 (telomeric repeat binding factor 2) is an essential component of the telomere-binding protein complex shelterin. TRF2 induces the formation of a special structure of telomeric DNA and counteracts activation of DNA damage-response pathways telomeres. TRF2 has a poorly characterized linker region (udTRF2) between its homodimerization and DNA-binding domains. Some lines of evidence have shown that this region could be involved in TRF2 interaction with nuclear lamina. Results: In this study, the fragment of the TERF2 gene encoding udTRF2 domain of telomere-binding protein TRF2 was produced by PCR and cloned into the pET32a vector. The resulting plasmid pET32a-udTRF2 was used for the expression of the recombinant udTRF2 in E. coli RosettaBlue (DE3). The protein was isolated and purified using ammonium sulfate precipitation followed by ion-exchange chromatography. The purified recombinant protein udTRF2 was injected into guinea pigs to generate polyclonal antibodies. The ability of anti-udTRF2 antibodies to bind endogenous TRF2 in human skin fibroblasts was tested by western blotting and immunofluorescent staining. Conclusions: In this study, the recombinant protein udTRF2 and antibodies to it were generated. Both protein and antibodies will provide a useful tool for investigation of the functions of the udTRF2 domain and its role in the interaction between TRF2 and nuclear lamina.
Assuntos
Animais , Cobaias , Proteína 2 de Ligação a Repetições Teloméricas/metabolismo , Anticorpos/metabolismo , Plasmídeos , Proteínas Recombinantes/metabolismo , Imuno-Histoquímica , Western Blotting , Cromossomos , Clonagem Molecular , Lâmina Nuclear , Proteína 2 de Ligação a Repetições Teloméricas/genética , Imunoprecipitação , Proteínas de Ligação a DNA/metabolismo , Escherichia coli/metabolismo , Anticorpos/isolamento & purificação , Formação de Anticorpos , NucleoproteínasRESUMO
Telomeric-repeat binding factor 2 (TRF2) as an important telomere binding protein, protects telomere ends and maintains the structural and functional stability and integrity of telomeres by interacting with five other essential core proteins (POT1, TRF1,TIN2, RAP1, and TPP1) to form a complex called shelterin. An increasing number of studies have discovered the abnormal expression of TRF2 in many tumors and that TRF2 is closely related to tumor occurrence, drug resistance, and tumor angiogenesis. Here, we review the latest research findings on the structure and function of TRF2, as well as the role of TRF2 in the occurrence, development,and treatment of tumors. We hope to provide novel perspectives for the prevention and treatment of tumors.