1. Top of page
  2. Abstract

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.


Y-box binding protein-1


embryonic stem


[3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt


phenazine ethosulfate

  • 1
    Wolffe AP, Tafuri S, Ranjan M, Familari M. The Y-box factors: a family of nucleic acid binding proteins conserved from Escherichia coli to man. New Biol 1992; 4: 2908.
  • 2
    Makino Y, Ohga T, Toh S, Koike K, Okumura K, Wada M, Kuwano M, Kohno K. Structural and functional analysis of the human Y-box binding protein (YB-1) gene promoter. Nucleic Acids Res 1996: 24: 18738.
  • 3
    Toh S, Nakamura T, Ohga T, Koike K, Uchiumi T, Wada M, Kuwano M, Kohno K. Genomic organization of the human Y-box protein (YB-1) gene. Gene 1998; 206: 937.
  • 4
    Didier DK, Schiffenbauer J, Woulfe SL, Zacheis M, Schwartz BD. Characterization of the cDNA encoding a protein binding to the major histocompatibility complex class II Y box. Proc Natl Acad Sci USA 1988; 85: 73226.
  • 5
    Ladomery M, Sommerville J. A role for Y-box proteins in cell proliferation. BioEssays 1995; 17: 911.
  • 6
    Wolffe AP. Structural and functional properties of the evolutionarily ancient Y-box family of nucleic acid binding proteins. BioEssays 1994; 16: 24551.
  • 7
    Uchiumi T, Kohno K, Tanimura H, Matsuo K, Sato S, Uchida Y, Kuwano M. Enhanced expression of the human multidrug resistance 1 gene in response to UV light irradiation. Cell Growth Differ 1993; 4: 14757.
  • 8
    Asakuno K, Kohno K, Uchiumi T, Kubo T, Sato S, Isono M, Kuwano M. Involvement of a DNA binding protein, MDR-NF1/YB-1, in human MDR1 gene expression by actinomycin D. Biochem Biophys Res Commun 1994; 199: 142835.
  • 9
    Ohga T, Uchiumi T, Makino Y, Koike K, Wada M, Kuwano M, Kohno K. Direct involvement of the Y-box binding protein YB-1 in genotoxic stress-induced activation of the human multidrug resistance 1 gene. J Biol Chem 1998; 273: 59976000.
  • 10
    Furukawa M, Uchiumi T, Nomoto M, Takano H, Morimoto RI, Naito S, Kuwano M, Kohno K. J Biol Chem 1998; 273: 105505.
  • 11
    Swamynathan SK, Nambiar A, Guntaka RV. Role of single-stranded DNA regions and Y-box proteins in transcriptional regulation of viral and cellular genes. FASEB J 1998; 2: 51522.
  • 12
    Matsumoto K, Wolffe AP. Gene regulation by Y-box proteins: coupling control of transcription and translation. Trends Cell Biol 1998; 8: 31823.
  • 13
    Sommerville J. Activities of cold-shock domain proteins in translation control. BioEssays 1999; 21: 31925.
  • 14
    Tafuri SR, Familari M, Wolffe AP. A mouse Y box protein, MSY1, is associated with paternal mRNA in spermatocytes. J Biol Chem 1993; 268: 1221320.
  • 15
    Ohga T, Koike K, No M, Makino Y, Itagaki Y, Tanimoto M, Kuwano M, Kohno, K. Role of the human Y box-binding protein YB-1 in cellular sensitivity to the DNA-damaging agents cisplatin, mitomycin C, and ultraviolet light. Cancer Res 1996; 56: 42248.
  • 16
    Koike K, Uchiumi T, Ohga T, Toh S, Wada M, Kohno K, Kuwano M. Nuclear translocation of the Y-box binding protein by ultraviolet irradiation. FEBS Lett 1997; 417: 3904.
  • 17
    Hasegawa SL, Doetsch PW, Hamilton KK, Martin AM, Okenquist SA, Lenz J, Boss JM. DNA binding properties of YB-1 and dbpA: binding to double-stranded, single-stranded, and abasic site containing DNAs. Nucleic Acids Res 1991; 19: 491520.
  • 18
    Ise T, Nagatani G, Imamura T, Kato K, Takano H, Nomoto M, Izumi H, Ohmori H, Okamoto T, Ohga T, Uchiumi T, Kuwano M, Kohno K. Transcription factor Y-box binding protein 1 binds preferentially to cisplatin-modified DNA and interacts with proliferating cell nuclear antigen. Cancer Res 1999; 59: 3426.
  • 19
    Hayakawa H, Uchiumi T, Fukuda T, Ashizuka M, Kohno K, Kuwano M, Sekiguchi M. Binding capacity of human YB-1 protein for RNA containing 8-oxoguanine. Biochemistry 2002; 41: 1273944.
  • 20
    Okamoto T, Izumi H, Imamura T, Takano H, Ise T, Uchiumi T, Kuwano M, Kohno K. Direct interaction of p53 with the Y-box binding protein, YB-1: a mechanism for regulation of human gene expression. Oncogene 2000; 19: 6194202.
  • 21
    Kamura T, Yahata H, Amada S, Ogawa S, Sonoda T, Kobayashi H, Mitsumoto M, Kohno K, Kuwano M, Nakano H. Is nuclear expression of Y box-binding protein-1 a new prognostic factor in ovarian serous adenocarcinoma Cancer 1999; 85: 24504.
  • 22
    Shibahara K, Sugio K, Osaki T, Uchiumi T, Maehara Y, Kohno K, Yasumoto K, Sugimachi K, Kuwano M. Nuclear expression of the Y-box binding protein, YB-1, as a novel marker of disease progression in non-small cell lung cancer. Clin Cancer Res 2001; 7: 31515.
  • 23
    Janz M, Harbeck N, Dettmar P, Berger U, Schmidt A, Jurchott K, Schmitt M, Royer HD. Y-box factor YB-1 predicts drug resistance and patient outcome in breast cancer independent of clinically relevant tumor biologic factors HER2, uPA and PAI-1. Int J Cancer 2001; 97: 27882.
  • 24
    Mansour SL, Thomas KR, Capecchi MR. Disruption of the proto-oncogene int-2 in mouse embryo-derived stem cells: a general strategy for targeting mutations to non-selectable genes. Nature 1988; 336: 34852.
  • 25
    Tsuzuki T, Fujii Y, Sakumi K, Tominaga Y, Nakao K, Sekiguchi M, Matsushiro A, Yoshimura Y, Morita T. Targeted disruption of the Rad51 gene leads to lethality in embryonic mice. Proc Natl Acad Sci USA 1996; 93: 623640.
  • 26
    Izumi H, Imamura T, Nagatani G, Ise T, Murakami T, Uramoto H, Torigoe T, Ishiguchi H, Yoshida Y, Nomoto M, Okamoto T, Uchiumi T, Kuwano M, Funa K, Kohno K. Y box-binding protein-1 binds preferentially to single-stranded nucleic acids and exhibits 3′[RIGHTWARDS ARROW]5′ exonuclease activity. Nucleic Acids Res 2001; 29: 12007.
  • 27
    Swamynathan S, Varma B, Weber K, Guntaka R. Targeted disruption of one allele of the Y-box protein gene, Chk-YB-1b, in DT40 cells results in major defects in cell cycle. Biochem Biophys Res Commun 2002; 296: 4517.
  • 28
    Oda Y, Sakamoto A, Shinohara N, Ohga T, Uchiumi T, Kohno K, Tsuneyoshi M, Kuwano M, Iwamoto Y. Nuclear expression of YB-1 protein correlates with P-glycoprotein expression in human osteosarcoma. Clin Cancer Res 1998; 4: 22737.
  • 29
    Shibao K, Takano H, Nakayama Y, Okazaki K, Nagata N, Izumi H, Uchiumi T, Kuwano M, Kohno K, Itoh H. Enhanced coexpression of YB-1 and DNA topoisomerase II alpha genes in human colorectal carcinomas. Int J Cancer 1999; 83: 7327.
  • 30
    Lorico A, Rappa G, Flavell RA, Sartorelli AC. Double knockout of the MRP gene leads to increased drug sensitivity in vitro. Cancer Res 1996; 56: 53515.
  • 31
    Tominaga Y, Tsuzuki T, Shiraishi A, Kawate H, Sekiguchi M. Alkylation-induced apoptosis of embryonic stem cells in which the gene for DNA-repair, methyltransferase, had been disrupted by gene targeting. Carcinogenesis 1997; 18: 88996.
  • 32
    Chu G. Cellular responses to cisplatin. The roles of DNA-binding proteins and DNA repair. J Biol Chem 1994; 269: 78790.
  • 33
    Stavrovskaya A. A cellular mechanisms of multidrug resistance of tumor cells. Biochemistry (Mosc) 2000; 65: 95106.
  • 34
    Zamble DB, Lippard SJ. Cisplatin and DNA repair in cancer chemotherapy. Trends Biochem Sci 1995; 20: 4359.
  • 35
    Timmer-Bosscha H, Mulder NH, de Vries EG. Modulation of cis-diamminedichloroplatinum(II) resistance: a review. Br J Cancer 1992; 66: 22738.
  • 36
    Arioka H, Nishio K, Ishida T, Fukumoto H, Fukuoka K, Nomoto T, Kurokawa H, Yokote H, Abe S, Saijo N. Enhancement of cisplatin sensitivity in high mobility group 2 cDNA-transfected human lung cancer cells. Jpn J Cancer Res 1999; 90: 10815.
  • 37
    He Q, Liang CH, Lippard SJ. Steroid hormones induce HMG1 overexpression and sensitize breast cancer cells to cisplatin and carboplatin. Proc Natl Acad Sci USA 2000; 97: 576872.
  • 38
    Brown SJ, Kellett PJ, Lippard SJ. Ixr1, a yeast protein that binds to platinated DNA and confers sensitivity to cisplatin. Science 1993; 261: 6035.
  • 39
    Damia G, Imperatori L, Stefanini M, D'Incalci M. Sensitivity of CHO mutant cell lines with specific defects in nucleotide excision repair to different anti-cancer agents. Int J Cancer 1996; 66: 77983.
  • 40
    De Silva IU, McHugh PJ, Clingen PH, Hartley JA. Defects in interstrand cross-link uncoupling do not account for the extreme sensitivity of ERCC1 and XPF cells to cisplatin. Nucleic Acids Res 2002; 30: 384856.
  • 41
    Uramoto H, Izumi H, Ise T, Tada M, Uchiumi T, Kuwano M, Yasumoto K, Funa K, Kohno K. p73 interacts with c-Myc to regulate Y-box-binding protein-1 expression. J Biol Chem 2002; 277: 3169472.