Screening of host proteins interacting with GP, VP30, NP,and L proteins of Ebola virus by yeast two-hybrid system / 军事医学

Objective To study the biological function of GP,VP30,NP and L proteins in Ebola virus(EBOV) infectious diseases,and to screen the host proteins which interact with GP,VP30,NP,and L proteins of EBOV from human liver cDNA library using yeast two-hybrid technique.Methods Recombinant PCR was used to construct bait plasmids pGBKT7-GP,pGBKT7-VP30,pGBKT7-NP and pGBKT7-L.Bait strains were combined with human liver cDNA library strains.The positive clones were analyzed by DNA sequencing and bioinformatics.A yeast recovery experiment was performed to further verify and exclude false positive results.Results We constructed bait plasmids pGBKT7-GP,pGBKT7-VP30,pGBKT7-NP and pGBKT7-L with the recombinant PCR method.Six host proteins which could interact with GP,VP30,NP,and L proteins were screened,including COMMD1,ALB,PSMD8,APOA2,CYP2E1,and HP.The yeast recovery experiment proved that COMMD1 and APOA2 might interact with NP protein.Conclusion A number of prey proteins which interact with GP,VP30,NP,and L proteins of EBOV are screened,which may provide reference for the research of EBOV.

Mutation Screening of Modifying Genes (Atox1 and Commd1) in Association with Wilson Disease.

Wilson disease (WD) results from accumulation of copper and is caused due to mutations in ATP7B, a copper transporting ATPase. Although WD is an established monogenic disorder, heterogeneity in phenotype is observed even among patients harboring mutations in ATP7B that would affect the mutant protein similarly. Such observations led to the speculation that there might be modifying loci that modulate the phenotype resulting from the aberration in the ATP7B gene. The expected genes coding for proteins that interact either directly with ATP7B or influence it indirectly might fit the role of modifier locus. ATOX1 and COMMD1 are the candidate genes that might play the role of a modifier locus having copper homeostasis pathway with such potential. To understand the role of modifying genes, we screened ATOX1 and COMMD1, a gene implicated in canine copper toxicosis, in 45 WD patients along with 50 healthy controls. This study did not yield satisfactory results concluding more patients to be analyzed. Keywords: Wilson Disease, ATOX1, COMMD1.

Expression and Localization of COMMD1 Proteins in Human Placentas from Women with Preeclampsia

PURPOSE: Recently, COMMD1 has been identified as a novel interactor and regulator of hypoxia-inducible factor-1 and nuclear factor kappa B transcriptional activity. The goal of this study was to determine the difference of COMMD1 expression in the placentas of women with normal and preeclamptic (PE) pregnancies. MATERIALS AND METHODS: Immnoperoxidase and immunofluorescent staining for COMMD1 was performed on nine normal and nine severe PE placental tissues, and COMMD1 mRNA expression was quantified by quantitative reverse transcription polymerase chain reaction. RESULTS: The expression of mRNA of COMMD1 was significantly higher in the study group than in the control group. The immunoreactivity was higher especially in the syncytiotrophoblast of PE placentas than in the control group. CONCLUSION: This study demonstrated increased placental COMMD1 expression in women with severe preeclampsia compared to that found in women with normal pregnancies, and this finding might contribute to a better understanding of the pathophysiology of preeclampsia.