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A Stable Upstream Stem-loop Structure Enhances Selection of the First 5'-ORF-AUG as a Main start codon for Translation Initiation of Human ACAT1 mRNA

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Acta Biochimica et Biophysica Sinica 2004年第4期36卷 259-268页

作者： Li YANG, Jiang CHEN, Catherine C. Y. CHANG1, Xin-Ying YANG, Zhen-Zhen WANG,Ta-Yuan CHANG1, and Bo-Liang LI* State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, the Chinese Academy of Sciences, Shanghai 200031, China Department of Biochemistry, Dartmouth Medical School, Hanover, NH 03756, USA 1 State Key Laboratory of Molecular Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences the Chinese Academy of SciencesShanghai 200031 China Department of Biochemistry Dartmouth Medical SchoolHanover NH 03756 USA

Human ACAT1 cDNA K1 was first cloned and functionally expressed in 1993. There are two adjacent in-frame AUG codons, AUG1397–1399 and AUG1415–1417, at 5'-terminus of the open reading frame (ORF, nt 1397–3049) of human ACAT1 mRNA corresponding to cDNA K1. In current work, these two adjacent in- frame AUGs at 5'-terminus of the predicted ORF (5'-ORF-AUGs) as start codons for translation initiation of human ACAT1 mRNA were characterized in detail. codon mutations indicated that both of these two adja- cent 5'-ORF-AUGs can be selected as start codons but the first 5'-ORF-AUG1397–1399 is a main start codon consistent with that of the predicted ORF of human ACAT1 mRNA. Further deletion and mutation analyses demonstrated that a stable upstream stem-loop structure enhanced the selection of the first 5'-ORF-AUG1397– as a main start codon, in addition to upstream nucleotide A in the –3 position, which is a key site of Kozak 1399 sequence. In addition, result of ACAT1 enzymatic activity assay showed no obvious difference between these two ACAT1 proteins respectively initiated from the two adjacent 5'-ORF-AUGs. This work showed that a stable upstream stem-loop structure could modulate the start codon selection during translation initiation of mRNAs that contain adjacent multi-5'-ORF-AUGs.

关键词： ACAT1 start codon Kozak sequence stem-loop structure

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The importance of start codon of nosM in nosiheptide production

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Chinese Journal of Natural Medicines 2015年第11期13卷 854-860页

作者： JIANG Lei XUE Yan-Jiu LIU Wei-Ying MA Min WU Xu-Ri WANG Shu-Zhen CHEN Yi-Jun State Key Laboratory of Natural Medicines and Laboratory of Chemical Biology China Pharmaceutical University

The present study was designed to investigate the effects of start codon of nosM on the biosynthesis of nosiheptide. Target genes were amplified by overlap PCR. After homologous recombination to construct engineered strains, nosiheptide production was analyzed by HPLC. Three mutants with different start codon ofnosMwere constructed, and nosihcptide production of each mutant was analyzed and compared. Replacement of the start eodon of nosM significantly decreased the production of nosiheptide. In conclusion, start codon usage could greatly affect the biosynthetic efficiency in the biosynthetic gene cluster of nosiheptide.

关键词： Nosiheptide start codon Leader peptide Biosynthesis

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Association of single nucleotide polymorphism of vitamin D receptor gene start codon and the suscepbility to prostate cancer in Han nationality in Hubei, China

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Asian Journal of Andrology 2004年第3期6卷 248-248页

作者： YiYang Shao-GangWang Zhang-QunYe Wei-MinYang DepartmentofUrology TongjiHospitalTongjiMedicalCollegeHuazhongUniversityofScienceandTechnologyWuhan430030China

Aim: To investigate the single nucleotide polymorphism of vitamin D receptor (VDR) gene start codon in the Han nationality in Hubei and its relationship to the susceptibility to prostate cancel (PCa). Methods: The VDR genotypes were determined by poly-merase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 80 patients with PCa and 96 normal male controls from the Han nationality in Hubei, using endonuclease Fok. Direct sequencing was done in part of the PCR products. Results: The frequency distribution of Fok I alleles in this cohort all followed the Hardy-Weinberg equilibrium. The distribution of genotypes and alleles had no significant difference between PCa patients and the controls (P>0.05). Conclusion: There was no significant relationship between Fok I polymorphism of VDR gene start codon and PCa in the Han nationality in Hubei.

关键词： prostate cancer vitamin D receptor gene start codon single nucleotide polymorphism Han nationality

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