title: The binding specificity of Translocated in LipoSarcoma/FUsed in Sarcoma with lncRNA transcribed from the promoter region of cyclin D1.
abstract: BACKGROUND
  Translocated in LipoSarcoma (TLS, also known as FUsed in Sarcoma) is an RNA/DNA binding protein whose mutation cause amyotrophic lateral sclerosis. In previous study, we demonstrated that TLS binds to long noncoding RNA, promoter-associated ncRNA-D (pncRNA-D), transcribed from the 5' upstream region of cyclin D1 (CCND1), and inhibits the expression of CCND1.
  
  
  RESULTS
  In order to elucidate the binding specificity between TLS and pncRNA-D, we divided pncRNA-D into seven fragments and examined the binding with full-length TLS, TLS-RGG2-zinc finger-RGG3, and TLS-RGG3 by RNA pull down assay. As a result, TLS was able to bind to all the seven fragments, but the fragments containing reported recognition motifs (GGUG and GGU) tend to bind more solidly. The full-length TLS and TLS-RGG2-zinc finger-RGG3 showed a similar interaction with pncRNA-D, but the binding specificity of TLS-RGG3 was lower compared to the full-length TLS and TLS-RGG2-zinc finger-RGG3. Mutation in GGUG and GGU motifs dramatically decreased the binding, and unexpectedly, we could only detect weak interaction with the RNA sequence with stem loop structure.
  
  
  CONCLUSIONS
  The binding of TLS and pncRNA-D was affected by the presence of GGUG and GGU sequences, and the C terminal domains of TLS function in the interaction with pncRNA-D.
authors.affiliation: Division of Gene Structure and Function, Research Center for Genomic Medicine, Saitama Medical University, 1397-1 Yamane, Hidaka-shi, Saitama 350-1241 Japan.
authors.forename: Ryoma
authors.initials: R
authors.lastname: Yoneda
authors.affiliation: Division of Gene Structure and Function, Research Center for Genomic Medicine, Saitama Medical University, 1397-1 Yamane, Hidaka-shi, Saitama 350-1241 Japan.
authors.forename: Shiho
authors.initials: S
authors.lastname: Suzuki
authors.affiliation: Institute of Advanced Energy, Kyoto University, Gokasho, Uji, Kyoto 611-0011 Japan ; Graduate School of Energy Science, Kyoto University, Gokasho, Uji, Kyoto 611-0011 Japan.
authors.forename: Tsukasa
authors.initials: T
authors.lastname: Mashima
authors.affiliation: Institute of Advanced Energy, Kyoto University, Gokasho, Uji, Kyoto 611-0011 Japan.
authors.forename: Keiko
authors.initials: K
authors.lastname: Kondo
authors.affiliation: Institute of Advanced Energy, Kyoto University, Gokasho, Uji, Kyoto 611-0011 Japan ; Graduate School of Energy Science, Kyoto University, Gokasho, Uji, Kyoto 611-0011 Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Nagata
authors.affiliation: Institute of Advanced Energy, Kyoto University, Gokasho, Uji, Kyoto 611-0011 Japan ; Graduate School of Energy Science, Kyoto University, Gokasho, Uji, Kyoto 611-0011 Japan.
authors.forename: Masato
authors.initials: M
authors.lastname: Katahira
authors.affiliation: Division of Gene Structure and Function, Research Center for Genomic Medicine, Saitama Medical University, 1397-1 Yamane, Hidaka-shi, Saitama 350-1241 Japan.
authors.forename: Riki
authors.initials: R
authors.lastname: Kurokawa
keywords: Long noncoding RNA; TLS/FUS; pncRNA
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