TY - JOUR
T1 - TET2 chemically modifies tRNAs and regulates tRNA fragment levels
AU - He, Chongsheng
AU - Bozler, Julianna
AU - Janssen, Kevin A.
AU - Wilusz, Jeremy E.
AU - Garcia, Benjamin A.
AU - Schorn, Andrea J.
AU - Bonasio, Roberto
N1 - Funding Information:
We thank R. Martienssen for support, encouragement, and discussions; M. Liu and R. Kohli for the kind gift of recombinant TET2 protein; T. Christopher for technical help; G. Xu for his generous gifts of Tet1/2/3-tKO cells; K. Ingvarsdottir and R. Warneford-Thomson for tissue culture help; and S. Erhardt for helpful discussion. R.B. acknowledges support from the NIH (R01GM127408). C.H. was supported in part by the National Natural Science Foundation of China (32070613, 31800687), the Natural Science Foundation of Hunan Province of China (2020JJ4179) and the Fundamental Research Funds for the Central Universities of China (531107051157). B.A.G. was supported in part by the NIH (R01GM110174, R01AI118891 and P01CA196539). A.J.S. acknowledges assistance from the Cold Spring Harbor Laboratory Shared Resources, which are funded in part by the Cancer Center Support Grant (5PP30CA045508). J.E.W. is a Rita Allen Foundation Scholar and is supported by NIH grant R35-GM119735.
Publisher Copyright:
© 2020, The Author(s), under exclusive licence to Springer Nature America, Inc.
PY - 2021/1
Y1 - 2021/1
N2 - The ten-eleven translocation 2 (TET2) protein, which oxidizes 5-methylcytosine in DNA, can also bind RNA; however, the targets and function of TET2–RNA interactions in vivo are not fully understood. Using stringent affinity tags introduced at the Tet2 locus, we purified and sequenced TET2-crosslinked RNAs from mouse embryonic stem cells (mESCs) and found a high enrichment for tRNAs. RNA immunoprecipitation with an antibody against 5-hydroxymethylcytosine (hm5C) recovered tRNAs that overlapped with those bound to TET2 in cells. Mass spectrometry (MS) analyses revealed that TET2 is necessary and sufficient for the deposition of the hm5C modification on tRNA. Tet2 knockout in mESCs affected the levels of several small noncoding RNAs originating from TET2-bound tRNAs that were enriched by hm5C immunoprecipitation. Thus, our results suggest a new function of TET2 in promoting the conversion of 5-methylcytosine to hm5C on tRNA and regulating the processing or stability of different classes of tRNA fragments.
AB - The ten-eleven translocation 2 (TET2) protein, which oxidizes 5-methylcytosine in DNA, can also bind RNA; however, the targets and function of TET2–RNA interactions in vivo are not fully understood. Using stringent affinity tags introduced at the Tet2 locus, we purified and sequenced TET2-crosslinked RNAs from mouse embryonic stem cells (mESCs) and found a high enrichment for tRNAs. RNA immunoprecipitation with an antibody against 5-hydroxymethylcytosine (hm5C) recovered tRNAs that overlapped with those bound to TET2 in cells. Mass spectrometry (MS) analyses revealed that TET2 is necessary and sufficient for the deposition of the hm5C modification on tRNA. Tet2 knockout in mESCs affected the levels of several small noncoding RNAs originating from TET2-bound tRNAs that were enriched by hm5C immunoprecipitation. Thus, our results suggest a new function of TET2 in promoting the conversion of 5-methylcytosine to hm5C on tRNA and regulating the processing or stability of different classes of tRNA fragments.
UR - http://www.scopus.com/inward/record.url?scp=85096447287&partnerID=8YFLogxK
U2 - 10.1038/s41594-020-00526-w
DO - 10.1038/s41594-020-00526-w
M3 - Article
C2 - 33230319
AN - SCOPUS:85096447287
VL - 28
SP - 62
EP - 70
JO - Nature Structural and Molecular Biology
JF - Nature Structural and Molecular Biology
SN - 1545-9993
IS - 1
ER -