Researcher Wang Enduo, who was elected as an academician of the Chinese Academy of Sciences in 2005, is mainly engaged in the quality control of protein biosynthesis. His research group is currently focusing on tRNA and related aminoacyl-tRNA synthetases, and has achieved many important results As far as this year is concerned, the research group of Academician Wang Enduo has published research findings in related research fields in PNAS, EMBO J, Nucleic Acids Res and other journals. The research team recently reported on the regulatory mechanism of preventing the incorporation of serine (Ser) into the threonine (Thr) codon during protein synthesis in eukaryotes.
Aminoacyl-tRNA synthetase (aaRS) is a key enzyme in the process of protein biosynthesis. It catalyzes the first step reaction in the process of protein biosynthesis-amino acylation of tRNA. The precise identification of tRNA by aminoacyl-tRNA synthetase ensures the accuracy of the transmission of genetic information from nucleic acid to protein. Therefore, studying the interaction between tRNA and aminoacyl-tRNA synthetase has important biological significance.
20 kinds of aaRS can be divided into 2 categories according to the structure and amino acylation site of tRNA, each of 10 kinds of aaRS, their catalytic reaction is the rate-limiting step of biosynthesis, quality control of protein synthesis directly affects the new peptide chain Function, which in turn affects the biological characteristics of the cell.
Threonyl-tRNA synthetase (ThrRS) belongs to the second type of aaRS, which is responsible for catalyzing the aminoacylation reaction between Thr and corresponding tRNAThr to generate threonyl-tRNA (Thr-tRNAThr), Thr-tRNAThr is further extended Transported to the ribosome to participate in protein biosynthesis. In this process, ThrRS must accurately select the corresponding amino acid and tRNA. However, Ser, a Thr analog in the cytoplasm, is easily misrecognized, activated, and aminoacylated by ThrRS. Therefore, ThrRS must remove the misactivated Ser through a hydrolysis reaction, which is called a redaction reaction. Aminoacylation and editing reactions have an important role in the normal physiology and metabolism of cells. Up to now, the understanding of the aminoacylation reaction catalyzed by the cytoplasmic ThrRS of eukaryotes and the mechanism of the editing reaction are at a blank stage.
In this article, the researchers systematically analyzed the eukaryotic cytoplasmic ThrRS and their evolutionary pathways, as well as the biological significance of the N-terminal extension of the eukaryotic ThrRS. The situation of ScThrRS recognizing Thr and other Thr analogs (Ser, Val, Cys, Gly, Ala) was analyzed. The mechanism of ScThrRS-catalyzed redaction reaction was clarified systematically, and each reaction path was quantified in the whole redaction reaction. The proportion and role.
In addition, through in-depth experiments, the researchers also identified key nucleotide residues that are critical to the editing reaction. They systematically studied the amino acid residues that performed key roles in each step of the reaction. The first haploid knockout strain of S. cerevisiae ScThrRS gene, through the in vivo genetic experiments to study the role of certain key amino acid residues in the aminoacylation reaction and editing reaction, also use this good platform to study different Cross-species cross-recognition of tRNAThr of Saccharomyces cerevisiae by species ThrRS
Through in vivo genetic experiments and in vitro proteomics analysis, they also found that even under very severe growth conditions, S. cerevisiae can still tolerate mutations of key amino acids in the editing domain of ThrRS, which implies yeast elongation factors and / or ribosomes The generated aminoacyl product Ser-tRNAThr can be further subjected to quality control in protein synthesis.
In another article published this year, Wang Enduo ’s research team also discovered for the first time that yeast uses “superwobbling†to read the leucine code, and they also used tl (uag) -Δ1-3 to get bands through site-directed mutation of genes. Yeast tRNALeu (UAG) variants with modified bases were used in vitro to study the function of tRNALeu variants screened in vivo, and the in vivo and in vitro data were consistent.
These studies not only enriched the understanding of the function of yeast tRNALeu, but also provided an effective research system for further studying the structure and function relationship of yeast tRNA.
About the Author:
Researcher Wang Enduo, academician of the Chinese Academy of Sciences, doctoral tutor, director of the Academic Committee of the State Key Laboratory of Molecular Biology
1965-1969 and 1978-1981, two graduate students from the Shanghai Institute of Biochemistry, Chinese Academy of Sciences. In 1986, he served as an associate researcher of the Institute of Biochemistry, and in 1993, as a researcher and doctoral tutor. 1984-1987 Visited at the University of California, Davis School of Medicine, October-December 1992, October-December 1994, January-April 1998 at the French National Science Research Center in Strasbourg, France Cooperative research of the Institute, research work in the Department of Biochemistry, Hong Kong University of Science and Technology, September 1996-February 1997, June-August 2000, September-October 2003, and July-September 2006 at Laval University, Canada The Department of Biochemistry conducts cooperative research. The eleventh National People's Congress, the tenth National Women's Federation Executive Committee. 1994 Shanghai "March 8" red flag bearer, 2000 Shanghai "March 8" red flag bearer, 1999-2000 Shanghai "March 8" red flag bearer, 1998-2000 Shanghai Model Worker, 2002 Shanghai Innovation Award The award is also named as the "3/8" red flag bearer in Shanghai, the Shanghai March 8 Women's Medal of Honor in 2005, and the National May 1st Labor Medal in 2007. Has hosted and undertaken 24 major and key research projects of the State and the Chinese Academy of Sciences. Published more than 100 papers and authorized 3 invention patents. The Shanghai Science and Technology Progress Award has 1 first prize and 2 second prizes, 1 National Natural Science Prize 2nd prize, the Chinese Academy of Sciences Excellent Graduate Instructor Award 5 times, and He Liang Heli Science and Technology Progress Award in 2006. More than 50 graduate students have won various awards including the National Outstanding Doctoral Dissertation Award and Nomination Award, the Chinese Academy of Sciences Outstanding Doctoral Dissertation Award and the Dean's Special Award. Director of the Academic Committee of the State Key Laboratory of Molecular Biology, member of the Academic Committee and Degree Evaluation Committee of the Institute, executive director of the Chinese Biochemistry and Molecular Biology, member of the American Society of Biochemistry and Molecular Biology. Executive deputy editor of "Life Science", member of the editorial board of "Journal of Biochemistry and Biophysics" and "China Science-C Series". Reviewer of "EMBO J", "Biochemistry (US)", "Nucleic Acids Research" and "J. Biol. Chem." He was elected to the Chinese Academy of Sciences in 2005, and was elected to the Third World Academy of Sciences in 2006.
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