TY - JOUR
T1 - APOBEC3B reporter myeloma cell lines identify DNA damage response pathways leading to APOBEC3B expression
AU - Yamazaki, Hiroyuki
AU - Shirakawa, Kotaro
AU - Matsumoto, Tadahiko
AU - Kazuma, Yasuhiro
AU - Matsui, Hiroyuki
AU - Horisawa, Yoshihito
AU - Stanford, Emani
AU - Sarca, Anamaria Daniela
AU - Shirakawa, Ryutaro
AU - Shindo, Keisuke
AU - Takaori-Kondo, Akifumi
N1 - Funding Information:
This work was partly supported by JSPS KAKENHI Grant numbers JP19H03502, 18H03992 for A.T-K., JP19K07591 for K.S. and by Amedisys Home Health and Hospice Care (AMED) under Grant Number JP19ck0106250, JP19cm0106501 for A.T-K. Research funding from Ono Pharmaceutical Co. for A.T-K. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.*%blankline%**%blankline%*
Publisher Copyright:
Copyright © 2020 Yamazaki et al.
PY - 2020/1/1
Y1 - 2020/1/1
N2 - Apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like (APOBEC) DNA cytosine deaminase 3B (A3B) is a DNA editing enzyme which induces genomic DNA mutations in multiple myeloma and in various other cancers. APOBEC family proteins are highly homologous so it is especially difficult to investigate the biology of specifically A3B in cancer cells. To easily and comprehensively investigate A3B function in myeloma cells, we used CRISPR/Cas9 to generate A3B reporter cells that contain 3×FLAG tag and IRES-EGFP sequences integrated at the end of the A3B gene. These reporter cells stably express 3×FLAG tagged A3B and the reporter EGFP and this expression is enhanced by known stimuli, such as PMA. Conversely, shRNA knockdown of A3B decreased EGFP fluorescence and 3×FLAG tagged A3B protein levels. We screened a series of anticancer treatments using these cell lines and identified that most conventional therapies, such as antimetabolites or radiation, exacerbated endogenous A3B expression, but recent molecular targeted therapeutics, including bortezomib, lenalidomide and elotuzumab, did not. Furthermore, chemical inhibition of ATM, ATR and DNA-PK suppressed EGFP expression upon treatment with antimetabolites. These results suggest that DNA damage triggers A3B expression through ATM, ATR and DNA-PK signaling.
AB - Apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like (APOBEC) DNA cytosine deaminase 3B (A3B) is a DNA editing enzyme which induces genomic DNA mutations in multiple myeloma and in various other cancers. APOBEC family proteins are highly homologous so it is especially difficult to investigate the biology of specifically A3B in cancer cells. To easily and comprehensively investigate A3B function in myeloma cells, we used CRISPR/Cas9 to generate A3B reporter cells that contain 3×FLAG tag and IRES-EGFP sequences integrated at the end of the A3B gene. These reporter cells stably express 3×FLAG tagged A3B and the reporter EGFP and this expression is enhanced by known stimuli, such as PMA. Conversely, shRNA knockdown of A3B decreased EGFP fluorescence and 3×FLAG tagged A3B protein levels. We screened a series of anticancer treatments using these cell lines and identified that most conventional therapies, such as antimetabolites or radiation, exacerbated endogenous A3B expression, but recent molecular targeted therapeutics, including bortezomib, lenalidomide and elotuzumab, did not. Furthermore, chemical inhibition of ATM, ATR and DNA-PK suppressed EGFP expression upon treatment with antimetabolites. These results suggest that DNA damage triggers A3B expression through ATM, ATR and DNA-PK signaling.
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U2 - 10.1371/journal.pone.0223463
DO - 10.1371/journal.pone.0223463
M3 - Article
C2 - 31914134
AN - SCOPUS:85077733644
VL - 15
JO - PLoS One
JF - PLoS One
SN - 1932-6203
IS - 1
M1 - e0223463
ER -