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Expression and Crystallization of Adenosine Deaminase Acting on tRNA
Author(s) -
Hurdle Carolyn M.,
Macbeth Mark R.
Publication year - 2020
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2020.34.s1.00640
Subject(s) - adar , inosine , adenosine deaminase , transfer rna , rna editing , rna , biochemistry , biology , adenosine , enzyme , gene , nucleotide , intron , chemistry , genetics
Adenosine Deaminase tRNA Specific 1 (ADAT1) is an enzyme in the adenosine deaminase acting on RNA (ADAR) family that causes site‐specific adenosine mutation in eukaryotic tRNA. tRNA deamination by ADAT1 changes the adenosine nucleotide to an inosine, impacting the control of specific genetic programs. These enzymes do not have double stranded RNA binding motifs like that of the ADARs but instead share the catalytic domain of the ADAR. While structures for several ADAR enzymes have been determined, there are no structures for ADAT1. This research seeks to determine the structure of the ADAT1 enzyme from C. albicans . The enzyme contains conserved residues that are proposed to bind Zn 2+ and the inositol hexakisphosphate co‐factor, yet it has truncations that are in non‐conserved regions when compared to ADAT1 genes from other organisms. The Candida gene has been subcloned and will be expressed in S. cerevisiae . Through crystallization and X‐ray diffraction, a structure will be determined and the mechanism of substrate recognition and catalysis will be evaluated. Support or Funding Information Holcomb Awards Committee Grant (MRM, 2019)