Studying nuclear functions of aminoacyl tRNA synthetases.
Aminoacyl tRNA synthetases (AARSs) are best known for their essential role in translation in the cytoplasm. The concept that AARSs also exist in the nucleus started to draw attention around the turn of the new millennium, when aminoacylated tRNAs were first found in the nuclei of Xenopus oocytes. It is now expected that all cytoplasmic AARSs are present in the nucleus. In addition to tRNA aminoacylation, nuclear AARSs were found to regulate a spectrum of biological processes and responses, with many AARSs functioning through regulation at the level of gene transcription. In this paper, we focus on describing methods that have been successfully implemented to study AARSs in transcriptional regulation. These include a cell fractionation assay to detect nuclear localization, an in vitro DNA-cellulose pull-down assay to determine DNA binding capacity, and a chromatin immunoprecipitation (ChIP)-DNA deep sequencing assay to identify DNA binding sites. Application of these methods would expand our understanding of AARS functions and reveal critical insights on the coordination of gene transcription and translation.
Departments of Chemical Physiology and Cell & Molecular Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA; The School of Medicine, Nankai University, 94 Weijin Road, Tianjin 300071, China. Electronic address: yishi@scripps.edu.
Yi
Y
Shi
Departments of Chemical Physiology and Cell & Molecular Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
Na
N
Wei
Departments of Chemical Physiology and Cell & Molecular Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA. Electronic address: xlyang@scripps.edu.
Xiang-Lei
XL
Yang
D000604:Amino Acyl-tRNA Synthetases / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D002458:Cell Fractionation / Q000379:methods; D002467:Cell Nucleus / Q000201:enzymology* / Q000235:genetics; D047369:Chromatin Immunoprecipitation / Q000379:methods*; D003600:Cytosol / Q000737:chemistry / Q000201:enzymology*; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D005455:Fluorescent Antibody Technique / Q000379:methods; D057809:HEK293 Cells; D006801:Humans; D011485:Protein Binding; D014176:Protein Biosynthesis; D021381:Protein Transport; D017422:Sequence Analysis, DNA; D014158:Transcription, Genetic
D004247:DNA; D000604:Amino Acyl-tRNA Synthetases
Aminoacyl tRNA synthetase; Chromatin immunoprecipitation; Nuclear function; Transcriptional regulation cell fractionation
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