Targeted disruption of one allele of the Y-box binding protein-1 (YB-1) gene in mouse embryonic stem cells and increased sensitivity to cisplatin and mitomycin C.
The eukaryotic Y-box binding protein-1 (YB-1) functions in various biological processes, including transcriptional and translational control, DNA repair, drug resistance, and cell proliferation. To elucidate the physiological role of the YB-1 protein, we disrupted one allele of mouse YB-1 in embryonic stem (ES) cells. Northern blot analysis revealed that YB-1(+/-) ES cells with one intact allele contain approximately one-half the amount of mRNA detected in wild-type (YB-1(+/+)) cells. We further found that the protein level of YB-1(+/-) cells was reduced to approximately 50-60% compared with that of YB-1(+/+) cells. However, no apparent growth difference was found between YB-1(+/-) and YB-1(+/+) cells. YB-1(+/-) cells showed increased sensitivity to cisplatin and mitomycin C, but not to etoposide, X-ray or UV irradiation, as compared to YB-1(+/+) cells. YB-1 may have the capacity to exert a protective role against cytotoxic effects of DNA damaging agents, and may be involved in certain aspects of drug resistance.
Shibahara
Kotaro
K
Department of Medical Biochemistry, Kyushu University, Fukuoka 812-8582, Japan.
Uchiumi
Takeshi
T
Fukuda
Takao
T
Kura
Shinobu
S
Tominaga
Yohei
Y
Maehara
Yoshihiko
Y
Kohno
Kimitoshi
K
Nakabeppu
Yusaku
Y
Tsuzuki
Teruhisa
T
Kuwano
Michihiko
M
D000483:Alleles; D000818:Animals; D018906:Antineoplastic Agents, Alkylating / Q000494:pharmacology*; D022762:CCAAT-Enhancer-Binding Proteins / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D002478:Cells, Cultured / Q000187:drug effects / Q000378:metabolism / Q000528:radiation effects; D002945:Cisplatin / Q000494:pharmacology*; D003114:Colony-Forming Units Assay; D004249:DNA Damage / Q000187:drug effects / Q000235:genetics / Q000528:radiation effects; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D004622:Embryo, Mammalian / Q000166:cytology; D005047:Etoposide / Q000494:pharmacology; D018390:Gene Targeting; D006579:Heterozygote; D051379:Mice; D016685:Mitomycin / Q000494:pharmacology*; D051839:NFI Transcription Factors; D012333:RNA, Messenger / Q000235:genetics; D013234:Stem Cells / Q000187:drug effects* / Q000378:metabolism / Q000528:radiation effects; D014157:Transcription Factors / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D014466:Ultraviolet Rays; D051840:Y-Box-Binding Protein 1
D018906:Antineoplastic Agents, Alkylating; D022762:CCAAT-Enhancer-Binding Proteins; D051839:NFI Transcription Factors; D012333:RNA, Messenger; D014157:Transcription Factors; D051840:Y-Box-Binding Protein 1; D016685:Mitomycin; D005047:Etoposide; D002945:Cisplatin