Effect of Biotinylation on the Structure and Function of a Yeast tRNA-Binding Protein

• Main Authors: Restu Nugraha, 尼古拉
• Other Authors: Chien-Chia Wang
• Format: Others
• Language: en_US
• Published: 2018
• Online Access: http://ndltd.ncl.edu.tw/handle/w6qtmj

碩士 === 國立中央大學 === 生命科學系 === 106 === Abstract Arc1p is a yeast-specific tRNA-binding protein that has ability to form a ternary complex with glutamyl-tRNA synthetase (GluRSc) and methionyl-tRNA synthetase (MetRS) in the cytoplasm. This complex can significantly enhance the aminoacylation efficiency of these two aaRSs to their respective cognate tRNAs. Recently, it was found that Arc1p can be biotinylated via post-translational modification at Lys86 (K86) in the N-domain. We herein studied the effect of K86 mutation on Arc1p’s structure and function. We found that mutation in K86R and K86A dramatically decreased the biotinylation level of Arc1p and altered its secondary structure. However, the mutant Arc1p could effectively rescue the cold-sensitive phenotype of an ARC1- strain, suggesting that biotinylation is dispensable for the rescue activity of Arc1p. Interestingly, K86R was more resistant to protease treatment than the wild-type, while K86A was more sensitive. Keywords: Arc1p, biotinylation, K86A, K86R, mutation.