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H X Liao

Publications and source records attributed to H X Liao.

14 recordsLinked to original sources
PubMed1991

Initiation of protein synthesis in animal mitochondria. Purification and characterization of translational initiation factor 2.

Bovine liver mitochondrial translational initiation factor 2 (IF-2mt) has been purified to near homogeneity. The scheme developed results in a 24,000-fold purification of the factor with about 26% recovery of activity. SDS-polyacrylamide gel electrophoresis indicates that IF-2mt has a subunit molecular mass of 85 kDa. IF-2mt promotes the binding of formyl(f)Met-tRNA to mitochondrial ribosomes but is inactive with the nonformylated derivative. IF-2mt is active on chloroplast 30 S ribosomal subunits, but IF-2chl has no activity in promoting fMet-tRNA binding to animal mitochondrial ribosomes. IF-2mt is sensitive to elevated temperatures and is inactivated by treatment with N-ethylmaleimide. It is partially protected from heat and N-ethylmaleimide inactivation by the presence of either GTP or GDP suggesting that guanine nucleotides may bind to this factor directly. The binding of fMet-tRNA to mitochondrial ribosomes requires the presence of GTP and is inhibited by GDP. DeoxyGTP is very effective in replacing GTP in promoting fMet-tRNA binding to ribosomes and some activity is also observed with ITP. No activity is observed with ATP, CTP, or UTP. Nonhydrolyzable analogs of GTP can promote formation of both 28 S and 55 S initiation complexes indicating that GTP hydrolysis is not required for subunit joining in the animal mitochondrial system.

Animals↗
PubMed1991

Interactions of bovine mitochondrial phenylalanyl-tRNA with ribosomes and elongation factors from mitochondria and bacteria.

A homologous in vitro poly(U)-directed translation system has been established using animal mitochondrial ribosomes, elongation factors (EF) and phenylalanyl-tRNA(Phe). The rate of incorporation of phenylalanine into polyphenylalanine in the mitochondrial system is slower than that observed for the homologous Escherichia coli system. E. coli ribosomes can be used in place of mitochondrial ribosomes in this system with only a slight decrease in the efficiency of phenylalanine incorporation from mitochondrial Phe-tRNA. However, E. coli elongation factor Tu (EF-Tu) cannot replace the mitochondrial EF-Tu in promoting the use of mitochondrial Phe-tRNA. The interaction between EF-Tu and mitochondrial Phe-tRNA was investigated by using the ability of EF-Tu to protect the aminoacyl-tRNA bond from hydrolysis. These results showed that both mitochondrial and E. coli EF-Tus are capable of interacting with mitochondrial Phe-tRNA. However, ribosomal A-site binding assays demonstrated that efficient binding of the mitochondrial Phe-tRNA to the ribosomal A-site was only obtained with the homologous mitochondrial EF-Tu.

Animals↗
PubMed1990

Identification and initial characterization of translational initiation factor 2 from bovine mitochondria.

The bovine liver mitochondrial factor that promotes the binding of fMet-tRNA to mitochondrial ribosomes, initiation factor 2 (IF-2mt), has been identified in the postribosomal supernatant fraction of isolated liver mitochondria. This factor has been purified approximately 5,000-fold and present preparations are estimated to be about 10% pure. IF-2mt has an apparent molecular weight of about 140,000 as determined by gel filtration chromatography. IF-2mt is active in stimulating fMet-tRNA binding to Escherichia coli ribosomes but E. coli IF-2 is not active in promoting initiator tRNA binding to animal mitochondrial ribosomes. The IF-2mt-mediated binding of fMet-tRNAi(Met) to mitochondrial ribosomes is dependent on the presence of a message such as poly(A,U,G) and on GTP. Nonhydrolyzable analogs of GTP are 2-3-fold less effective in promoting initiation complex formation on mitochondrial ribosomes than is GTP suggesting that IF-2mt is capable of recycling to some extent under the current assay conditions.

Animals↗
PubMed1990

Effects of length and mRNA secondary structure on the interaction of bovine mitochondrial ribosomes with messenger RNA.

The mRNA for cytochrome oxidase subunit II (CoII) from bovine mitochondria binds to the small subunit of the mitochondrial ribosome in the absence of auxiliary factors. The synthetic polymer poly(U) is effective in competing with CoII mRNA for binding, although the polymer poly(A,U,G) competes very weakly. The effects of mRNA length on the interaction between the 28 S ribosomal subunit and mRNA have been examined using truncated derivatives of CoII mRNA. These results indicate that there is a minimum length of approximately 400 nucleotides required for the efficient binding of the mRNA to the small subunit. Shorter mRNAs will bind, but do so with much lower association constants. mRNAs of various lengths but with reduced secondary structure were prepared by substituting ITP for GTP during in vitro transcription reactions. These derivatives show the same effects of length as do the normal mRNA, indicating that RNA secondary structure is not a critical factor in subunit-mRNA interaction. The binding of the mRNA to the 28 S subunit is not influenced by the presence of guanine nucleotides or by the presence of a triphosphate at the 5' end of the RNA.

Animals↗
PubMed1989

Interaction of bovine mitochondrial ribosomes with messenger RNA.

The gene for subunit II of cytochrome oxidase (CoII) from bovine mitochondria has been cloned behind a T7 promoter and the corresponding mRNA synthesized in vitro. The RNA transcribed from this vector has a single nucleotide 5' to the start AUG and, thus, corresponds closely to the native mRNA. It binds to the small 28 S ribosomal subunit of bovine mitochondria but not to the large (39 S) subunit or to 55 S ribosomes. The binding occurs readily in the absence of auxiliary initiation factors or initiator tRNA. The complex formed appears to contain 1 mRNA/28 S subunit. The observed binding is specific for mRNA since neither tRNA nor ribosomal RNA can act as competitive inhibitors. The interaction of the mRNA with the 28 S subunit does not require an AUG codon near the 5' end and constructs containing 5' leaders of more than 100 nucleotides still bind efficiently. About 5% of the bound mRNA is protected from digestion by T1 RNase. The protected fragments do not arise from a specific region of the mRNA since they hybridize to several restriction fragments of the cloned CoII gene.

Animals↗
PubMed1984

Antigenic difference between viral strains causing classical and mild types of epidemic hemorrhagic fever with renal syndrome in China.

The antigenic relationship between viral isolates from Apodemus and Rattus that appear to cause the classical and mild types of epidemic hemorrhagic fever (EHF) in China was studied by cross-immunofluorescence, cross-neutralization, immunofluorescence blocking tests, and cross-enzyme-linked immunosorbent assay (ELISA). Obvious antigenic diversity between the isolates was demonstrated by cross-neutralization, immunofluorescence blocking tests, and cross-ELISA. Antisera from patients with classical EHF neutralized viruses of both types to a similar degree, but antisera from patients with mild EHF showed little neutralization of apodemus virus. Similarly, antisera from classical EHF blocked immunofluorescence by monoclonal antibody (25-1 McAb) derived from apodemus virus to both viral antigens, but antisera from mild EHF gave only low-grade blocking against apodemus viral antigen. Direct antigenic titrations of both viral strains by cross-ELISA yielded similar results. That distinct antigenic differences exist between viral strains causing these two types of EHF might be of great importance to the serological differentiation of the viruses and the study of EHF vaccine.

Animals↗
PubMed1983

Etiologic studies of epidemic hemorrhagic fever (hemorrhagic fever with renal syndrome).

Two strains of epidemic hemorrhagic fever (EHF) virus were isolated from the lung tissues of Apodemus agrarius mice that were captured in an area where EHF is endemic. The strains were isolated by passages in A. agrarius mice from a nonendemic area. Identification of the isolates by usual procedures was confirmed by repeated blind tests with coded sera. Contamination with certain known viruses such as reovirus, adenovirus (types 3 and 7), and other pathogens, such as murine typhus rickettsiae and Leptospira, which may be naturally present in wild rodents, appeared to have been ruled out. The antigen slides made from these isolates are in use in the specific diagnosis and seroepidemiologic studies of EHF. The first successful application is the serodiagnosis of a mild type of hemorrhagic fever that occurs with characteristic epidemiologic features in certain provinces of China.

Animals↗