Mapping of homologous interaction sites in the hepatitis B virus core protein.
نویسندگان
چکیده
Hepatitis B virus consists of an outer envelope and an inner capsid, or core, that wraps around the small genome plus the viral replication enzyme. The icosahedrally symmetric nucleocapsid is assembled from multiple dimeric subunits of a single 183-residue capsid protein, which must therefore contain interfaces for monomer dimerization and for dimer multimerization. The atomic structure of the protein is not known, but electron microscopy-based image reconstructions suggested a hammerhead shape for the dimer and, very recently, led to a tentative model for the main chain trace. Here we used a combination of interaction screening techniques and functional analyses of core protein variants to define, at the primary sequence level, the regions that mediate capsid assembly. Both the two-hybrid system and the pepscan technique identified a strongly interacting region I between amino acids (aa) 78 and 117 that probably forms part of the dimer interface. Surprisingly, mutations in this region, in the context of a C-terminally truncated but assembly-competent core protein variant, had no detectable effect on assembly. By contrast, mutations in a second region, bordered by aa 113 and 143, markedly influenced capsid stability, strongly suggesting that this region II is the main contributor to dimer multimerization. Based on the electron microscopic data, it must therefore be located at the basal tips of the dimer, experimentally supporting the proposed main chain trace.
منابع مشابه
Expression of Hepatitis B Virus Core Antigen in Native and Fusion Forms in E. coli
DNA coding for the core antigen from hepatitis B Virus (HBcAg) was amplified, cloned and propagated in E. coli. The core protein was expressed in E. coli and the product was readily detected by Western blot. This protein can be used as a diagnostic material in serum screening tests. To increase the level of expression of this antigen in bacteria, two plasmids were constructed in which the gene ...
متن کاملOverexpression of Full-Length Core Protein of Hepatitis C Virus by Escherichia coli Cultivated in Stirred Tank Fermentor
The mature core protein of the Hepatitis C virus (HCVC173) carrying pelB as a signal peptide (PelB::core) was overexpressed in Escherichia coli as 18% and 23.3% of the host’s total protein, in flask and fermentor cultivation, respectively. A final specific yield of 25 ± 1 mg HCVC173/g dry cell weight and an overallproductivity of 51±1 mg HCVC173/l/h were obtained in the stirred-tank ferme...
متن کاملMutations at Nucleotide 1762, 1764 and 1766 of Hepatitis B Virus X Gene in Patients with Chronic Hepatitis B and Hepatitis B-Related Cirrhosis
Abstract Background and objective: Hepatitis B virus (HBV) is a DNA virus with high tendency toward hepatic tissue. There are currently about 3 million HBV-infected people and 350 to 400 million chronic carriers of this virus in the world. X protein plays a role in the over-expression of oncogenes, carcinogenicity of liver cells and overlaps with the basal co...
متن کاملPriming Hepatitis B Surface (HBsAg)- and Core Antigen (HBcAg)-Specific Immune Responses by Chimeric, HBcAg with a HBsAg ‘a’ Determinant
We developed an immunogen to stimulate multivalent immunity against hepatitis B surface antigen (HBsAg) and hepatitis B core antigens (HBcAg). Immune responses specific for both HBsAg and HBcAg play an important role in controlling the infection. HBsAg-specific antibodies mediate elimination of virions at an early stage of infection and prevent the spread of virus. The immunogen was constructed...
متن کاملCpG Motif as an Adjuvant in Immunization of a Recombinant Plasmid Encoding Hepatitis C Virus Core Protein
متن کامل
Truncated Hepatitis B virus like nanoparticles: A novel drug delivery platform for cancer therapy
Nowadays, Nano-sized drug delivery systems have been studied extensively for theirpotential in cancer therapy. Various drug nanocarriers are being developed including liposomes, micelles, and Virus like nanoparticles (VLNPs). VLNPs offer many advantages for developing smart drug delivery systems due to their precise and repeated structures and relatively large cargo capacities. Truncated ...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Journal of virology
دوره 72 6 شماره
صفحات -
تاریخ انتشار 1998