TY - JOUR
T1 - Specific binding of PCBP1 to heavily oxidized RNA to induce cell death
AU - Ishii, Takashi
AU - Hayakawa, Hiroshi
AU - Igawa, Tatsuhiro
AU - Sekiguchi, Takeshi
AU - Sekiguchi, Mutsuo
N1 - Funding Information:
We thank Masumi Hidaka, Ryosuke Fujikane, and Brian Quinn for useful comments and Hisako Takeuchi for her excellent technical assistance. This work was supported by grants from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (MEXT), including the MEXT-supported Program for the Strategic Research Foundation at Private Universities Grant S1411042 and Grants in-Aid for Scientific Research 16K07260, 22370003, and 26340040.
Funding Information:
ACKNOWLEDGMENTS. We thank Masumi Hidaka, Ryosuke Fujikane, and Brian Quinn for useful comments and Hisako Takeuchi for her excellent technical assistance. This work was supported by grants from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (MEXT), including the MEXT-supported Program for the Strategic Research Foundation at Private Universities Grant S1411042 and Grants in-Aid for Scientific Research 16K07260, 22370003, and 26340040.
Publisher Copyright:
© 2018 National Academy of Sciences. All Rights Reserved.
PY - 2018/6/26
Y1 - 2018/6/26
N2 - In aerobically growing cells, the guanine base of RNA is oxidized to 8-oxo-7,8-dihydroguanine (8-oxoG), which induces alteration in their gene expression. We previously demonstrated that the human AUF1 protein binds to 8-oxoG in RNA to induce the selective degradation of oxidized messenger RNA. We herein report that the poly(C)-binding protein PCBP1 binds to more severely oxidized RNA to activate apoptosis-related reactions. While AUF1 binds to oligoribonucleotides carrying a single 8-oxoG, PCBP1 does not bind to such oligoribonucleotides but instead binds firmly to oligoribonucleotides in which two 8-oxoG residues are located nearby. PCBP1-deficient cells, constructed from the human HeLa S3 line using the CRISPR-Cas9 system, exhibited higher survival rates than HeLa S3 cells when small doses of hydrogen peroxide were applied. The levels of caspase-3 activation and PARP-1 cleavage in the PCBP1-deficient cells were significantly lower than those in wild-type cells. The structure–function relationship of PCBP1 was established with the use of PCBP1 mutant proteins in which the conserved KH domains were defective. Human cells appear to possess two distinct mechanisms, one controlled by AUF1 and the other by PCBP1, with the former functioning when messenger RNA is moderately oxidized and the latter operating when the RNA is more severely damaged.
AB - In aerobically growing cells, the guanine base of RNA is oxidized to 8-oxo-7,8-dihydroguanine (8-oxoG), which induces alteration in their gene expression. We previously demonstrated that the human AUF1 protein binds to 8-oxoG in RNA to induce the selective degradation of oxidized messenger RNA. We herein report that the poly(C)-binding protein PCBP1 binds to more severely oxidized RNA to activate apoptosis-related reactions. While AUF1 binds to oligoribonucleotides carrying a single 8-oxoG, PCBP1 does not bind to such oligoribonucleotides but instead binds firmly to oligoribonucleotides in which two 8-oxoG residues are located nearby. PCBP1-deficient cells, constructed from the human HeLa S3 line using the CRISPR-Cas9 system, exhibited higher survival rates than HeLa S3 cells when small doses of hydrogen peroxide were applied. The levels of caspase-3 activation and PARP-1 cleavage in the PCBP1-deficient cells were significantly lower than those in wild-type cells. The structure–function relationship of PCBP1 was established with the use of PCBP1 mutant proteins in which the conserved KH domains were defective. Human cells appear to possess two distinct mechanisms, one controlled by AUF1 and the other by PCBP1, with the former functioning when messenger RNA is moderately oxidized and the latter operating when the RNA is more severely damaged.
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U2 - 10.1073/pnas.1806912115
DO - 10.1073/pnas.1806912115
M3 - Article
C2 - 29891675
AN - SCOPUS:85049054601
VL - 115
SP - 6715
EP - 6720
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 26
ER -