Tpr-like Protein Biological Function And Mechanism Of Action

Objective:To investigate NYD-SP14 protein and those proteins whichcan interact with NYD-SP14,further explore the effect of NYD-SP14 on cell division growth and the mechanism of these effects on the spermatogenesis and tumorigeness.Methods:A series of yeast expressing vector and Fluorescence expressing vector of NYD-SP14,PP2A,HRB2,HSPT0,HSP90 were constructed by using gene recombination technique.The expression profile of NYD-SP14 was viewed through RT-PCR.Yeast two-hybrid and GST pull-down assays revealed which proteins interact with NYD-SP14 and which region of these proteins is essential.Then we can suppose NYD-SP14'S and these interactives's possible function.Express,purificat and obtain NYD-SP14 protein.Then we further use NYD-SP14 protein to immune Babla/C mouse to follow resches.GFP-NYD-SP 14,GFP-HRB2 steady expression HELA cell was constructed,then the subcellular location of GFP-NYD-SP14,GFP-HRB2 fusion proteins was observed.Results:The expression profile of NYD-SP14 showed that NYD-SP14 is highly expressed in testis-specific and cancer tissue,indicating a potential role for NYD-SP14 in testis and cancer development and spermatogenesi-s. Yeast two-hybrid and GST pull-down assays revealed NYD-SP14 can intract with PP2R1A,PP2AC,HRB2,HSP70,HSP90 and otherwise NYD-SP 14's 272～387aa region,PP2R1A's 183～463aa.And TPR domain in NYD-SP14 and C terminatio EEVD of HSP70 and HSP90 are also testified.NYD-SP14 fusion protein was expressed in the cytoplasm, while HRB2 was expressed in the cytoplasm before cell division,but after cell division it was show in the nucleus.Conclusion:Bioinformatic analysis indicates that NYD-SP14 protein is a member of TPR domain family.Evolutional analysis reveals that the structural domain in the N-terminus TPR domain originates and three continuous TPR domain. Therefore,NYD-SP14 conforms to the common features of TPR homology proteins.The results NYD-SP14 may have the common features of TPR homology proteins which could induce cell proliferation. NYD-SP14 was highly expressed in testis and cancer tissue.This result further suggested NYD-SP14 may be a cancer-testis gene and take part not in only cell proliferation but also in canser and testis.This study lays a foundation for the functional study of NYD-SP14,and provides new pathway for the clinical application of gene therapies for canser and testis. Objective:To investigate interaction between 64 different kinds of TPR domain homology proteins which contain three consecutive TPR domains and PP2R1A,PP2AC,HSP70,HSP90,further explore the effect of TPR domain homology proteins phosphorylation on cell division growth and the mechanism of these interactions.Methods:constructing 52 kinds of TPR domain homology proteins which contain three consecutive TPR domains yeast expressing vector.Yeast two-hybrid and non-S³⁵GST pull-down assays revealed if 64 kinds of TPR domain homology proteins which contain three consecutive TPR domains can interact with PP2R1A,PP2AC,HSP70,HSP90 and which region of these proteins is essential.Results:A series of Yeast two-hybrid and non-S³⁵GST pull-down assays revealed 64 kinds of TPR domain homology proteins which contain three consecutive TPR domains can interact with PP2R1A,PP2AC,HSP70,HSP90 and three consecutive TPR domains region of these proteins is essential to these interactions.Conclusion:We find 52 new kinds of interactions between TPR domain proteins and PP2R1A,PP2AC,HSP70,HSP90.And these interactions by their TPR domain is essential to these interactions.So we suggest it is universal function that TPR domain proteins which contain three consecutive TPR domains.Then these founds offer a novel mode to study new proteins interactions between TPR domain proteins and PP2R1A,HSP70,HSP90.