Interleukin Enhancer-binding Factor 3/NF110 Is a Target of YM155, a Suppressant of Survivin

Naoto Nakamura,Tomohiro Yamauchi,Masashi Hiramoto,Masatoshi Yuri,Masanori Naito,Masahiro Takeuchi,Kentaro Yamanaka,Aya Kita,Takahito Nakahara,Isao Kinoyama,Akira Matsuhisa,Naoki Kaneko,Hiroshi Koutoku,Masao Sasamata,Hiroyuki Yokota,Shigeki Kawabata,Kiyoshi Furuichi

doi:10.1074/mcp.m111.013243

Naoto Nakamura, Tomohiro Yamauchi + Show 15 more

Open Access

https://doi.org/10.1074/mcp.m111.013243

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Abstract

Survivin is responsible for cancer progression and drug resistance in many types of cancer. YM155 selectively suppresses the expression of survivin and induces apoptosis in cancer cells in vitro and in vivo. However, the mechanism underlying these effects of YM155 is unknown. Here, we show that a transcription factor, interleukin enhancer-binding factor 3 (ILF3)/NF110, is a direct binding target of YM155. The enhanced survivin promoter activity by overexpression of ILF3/NF110 was attenuated by YM155 in a concentration-dependent manner, suggesting that ILF3/NF110 is the physiological target through which YM155 mediates survivin suppression. The results also show that the unique C-terminal region of ILF3/NF110 is important for promoting survivin expression and for high affinity binding to YM155.

Highlights

Survivin is a member of the inhibitor of apoptosis protein family and intersects the multiple pathways involved in tumor proliferation and inhibition of apoptosis, as a nodal molecule [1,2,3]
The existence of the three molecules in the nucleus was confirmed in an input sample used for our affinity purification, only interleukin enhancer-binding factor 3 (ILF3)/NF110 and ILF2/NF45 were purified from the nuclear extract by YM155 beads (Fig. 2b)
It is known that ILF2 forms complexes with both ILF3/NF110 and ILF3/NF90 [24], so if ILF2 bound directly to YM155, both ILF3/NF90 and ILF3/NF110 would have been detected via this indirect association

Summary

Introduction

Survivin is a member of the inhibitor of apoptosis protein family and intersects the multiple pathways involved in tumor proliferation and inhibition of apoptosis, as a nodal molecule [1,2,3]. Construction of Expression Vectors—To amplify the ILF31 isoforms (ILF3–1a, -1b, -2a, -2b, -3a, and -3b) and deletion mutants (ILF3-⌬N and ILF3-⌬C), PCR was performed using human testis MarathonReady cDNA (Clontech) with the primers listed in supplemental Table 2. Pulldown Assay—The pulldown assays were performed using lysates of PC-3 cells overexpressing ILF3 isoforms or deletion mutants, all of which contained a C-terminal FLAG epitope tag.

Results

Conclusion