TY - JOUR
T1 - Queuine Salvaging in the Human Parasite Entamoeba histolytica
AU - Sarid, Lotem
AU - Sun, Jingjing
AU - Chittrakanwong, Jurairat
AU - Trebicz-Geffen, Meirav
AU - Ye, Jun
AU - Dedon, Peter C.
AU - Ankri, Serge
N1 - Publisher Copyright: © 2022 by the authors.
PY - 2022/8
Y1 - 2022/8
N2 - Queuosine (Q) is a naturally occurring modified nucleoside that occurs in the first position of transfer RNA anticodons such as Asp, Asn, His, and Tyr. As eukaryotes lack pathways to synthesize queuine, the Q nucleobase, they must obtain it from their diet or gut microbiota. Previously, we described the effects of queuine on the physiology of the eukaryotic parasite Entamoeba histolytica and characterized the enzyme EhTGT responsible for queuine incorporation into tRNA. At present, it is unknown how E. histolytica salvages queuine from gut bacteria. We used liquid chromatography–mass spectrometry (LC–MS) and N-acryloyl-3-aminophenylboronic acid (APB) PAGE analysis to demonstrate that E. histolytica trophozoites can salvage queuine from Q or E. coli K12 but not from the modified E. coli QueC strain, which cannot produce queuine. We then examined the role of EhDUF2419, a protein with homology to DNA glycosylase, as a queuine salvage enzyme in E. histolytica. We found that glutathione S-transferase (GST)-EhDUF2419 catalyzed the conversion of Q into queuine. Trophozoites silenced for EhDUF2419 expression are impaired in their ability to form Q-tRNA from Q or from E. coli. We also observed that Q or E. coli K12 partially protects control trophozoites from oxidative stress (OS), but not siEhDUF2419 trophozoites. Overall, our data reveal that EhDUF2419 is central for the direct salvaging of queuine from bacteria and for the resistance of the parasite to OS.
AB - Queuosine (Q) is a naturally occurring modified nucleoside that occurs in the first position of transfer RNA anticodons such as Asp, Asn, His, and Tyr. As eukaryotes lack pathways to synthesize queuine, the Q nucleobase, they must obtain it from their diet or gut microbiota. Previously, we described the effects of queuine on the physiology of the eukaryotic parasite Entamoeba histolytica and characterized the enzyme EhTGT responsible for queuine incorporation into tRNA. At present, it is unknown how E. histolytica salvages queuine from gut bacteria. We used liquid chromatography–mass spectrometry (LC–MS) and N-acryloyl-3-aminophenylboronic acid (APB) PAGE analysis to demonstrate that E. histolytica trophozoites can salvage queuine from Q or E. coli K12 but not from the modified E. coli QueC strain, which cannot produce queuine. We then examined the role of EhDUF2419, a protein with homology to DNA glycosylase, as a queuine salvage enzyme in E. histolytica. We found that glutathione S-transferase (GST)-EhDUF2419 catalyzed the conversion of Q into queuine. Trophozoites silenced for EhDUF2419 expression are impaired in their ability to form Q-tRNA from Q or from E. coli. We also observed that Q or E. coli K12 partially protects control trophozoites from oxidative stress (OS), but not siEhDUF2419 trophozoites. Overall, our data reveal that EhDUF2419 is central for the direct salvaging of queuine from bacteria and for the resistance of the parasite to OS.
KW - Animals
KW - Entamoeba histolytica
KW - Entamoeba histolytica/metabolism
KW - Escherichia coli/metabolism
KW - Guanine/analogs & derivatives
KW - Humans
KW - Parasites/metabolism
KW - RNA, Transfer/genetics
KW - oxidative stress resistance
KW - queuine
KW - queuosine
KW - tRNA modifications
UR - http://www.scopus.com/inward/record.url?scp=85136671833&partnerID=8YFLogxK
U2 - https://doi.org/10.3390/cells11162509
DO - https://doi.org/10.3390/cells11162509
M3 - مقالة
C2 - 36010587
SN - 2073-4409
VL - 11
JO - Cells
JF - Cells
IS - 16
M1 - 2509
ER -