Analysis of transforming activity of human synovial sarcoma-associated chimeric protein SYT-SSX1 bound to chromatin remodeling factor hBRM/hSNF2α

Makoto Nagai, Shinya Tanaka, Masumi Tsuda, Shuichi Endo, Hiroyuki Kato, Hiroshi Sonobe, Akio Minami, Hiroaki Hiraga, Hiroshi Nishihara, Hirofumi Sawa, Kazuo Nagashima

Research output: Contribution to journalArticle

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Abstract

Human synovial sarcoma has been shown to exclusively harbor the chromosomal translocation t(X;18) that produces the chimeric gene SYT-SSX. However, the role of SYT-SSX in cellular transformation remains unclear. In this study, we have established 3Y1 rat fibroblast cell lines that constitutively express SYT, SSX1, and SYT-SSX1 and found that SYT-SSX1 promoted growth rate in culture, anchorage-independent growth in soft agar, and tumor formation in nude mice. Deletion of the N-terminal 181 amino acids of SYT-SSX1 caused loss of its transforming activity. Furthermore, association of SYT-SSX1 with the chromatin remodeling factor hBRM/hSNF2α, which regulates transcription, was demonstrated in both SYT-SSX1-expressing 3Y1 cells and in the human synovial sarcoma cell line HS-SY-II. The binding region between the two molecules was shown to reside within the N-terminal 181 amino acids stretch (aa 1-181) of SYT-SSX1 and 50 amino acids (aa 156-205) of hBRM/hSNF2α and we found that the overexpression of this binding region of hBRM/hSNF2α significantly suppressed the anchorage-independent growth of SYT-SSX1-expressing 3Y1 cells. To analyze the transcriptional regulation by SYT-SSX1, we established conditional expression system of SYT-SSX1 and examined the gene expression profiles. The down-regulation of potential tumor suppressor DCC was observed among 1,176 genes analyzed by microarray analysis, and semi-quantitative reverse transcription-PCR confirmed this finding. These data clearly demonstrate transforming activity of human oncogene SYT-SSX1 and also involvement of chromatin remodeling factor hBRM/hSNF2α in human cancer.

Original languageEnglish
Pages (from-to)3843-3848
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume98
Issue number7
DOIs
Publication statusPublished - 2001 Mar 27
Externally publishedYes

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Synovial Sarcoma
Chromatin Assembly and Disassembly
Human Activities
Proteins
Amino Acids
SYT-SSX fusion protein
Growth
Cell Line
Neoplasms
Genetic Translocation
Microarray Analysis
Oncogenes
Transcriptome
Nude Mice
Genes
Reverse Transcription
Agar
Down-Regulation
Fibroblasts

ASJC Scopus subject areas

  • General

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Analysis of transforming activity of human synovial sarcoma-associated chimeric protein SYT-SSX1 bound to chromatin remodeling factor hBRM/hSNF2α. / Nagai, Makoto; Tanaka, Shinya; Tsuda, Masumi; Endo, Shuichi; Kato, Hiroyuki; Sonobe, Hiroshi; Minami, Akio; Hiraga, Hiroaki; Nishihara, Hiroshi; Sawa, Hirofumi; Nagashima, Kazuo.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 98, No. 7, 27.03.2001, p. 3843-3848.

Research output: Contribution to journalArticle

Nagai, Makoto ; Tanaka, Shinya ; Tsuda, Masumi ; Endo, Shuichi ; Kato, Hiroyuki ; Sonobe, Hiroshi ; Minami, Akio ; Hiraga, Hiroaki ; Nishihara, Hiroshi ; Sawa, Hirofumi ; Nagashima, Kazuo. / Analysis of transforming activity of human synovial sarcoma-associated chimeric protein SYT-SSX1 bound to chromatin remodeling factor hBRM/hSNF2α. In: Proceedings of the National Academy of Sciences of the United States of America. 2001 ; Vol. 98, No. 7. pp. 3843-3848.
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abstract = "Human synovial sarcoma has been shown to exclusively harbor the chromosomal translocation t(X;18) that produces the chimeric gene SYT-SSX. However, the role of SYT-SSX in cellular transformation remains unclear. In this study, we have established 3Y1 rat fibroblast cell lines that constitutively express SYT, SSX1, and SYT-SSX1 and found that SYT-SSX1 promoted growth rate in culture, anchorage-independent growth in soft agar, and tumor formation in nude mice. Deletion of the N-terminal 181 amino acids of SYT-SSX1 caused loss of its transforming activity. Furthermore, association of SYT-SSX1 with the chromatin remodeling factor hBRM/hSNF2α, which regulates transcription, was demonstrated in both SYT-SSX1-expressing 3Y1 cells and in the human synovial sarcoma cell line HS-SY-II. The binding region between the two molecules was shown to reside within the N-terminal 181 amino acids stretch (aa 1-181) of SYT-SSX1 and 50 amino acids (aa 156-205) of hBRM/hSNF2α and we found that the overexpression of this binding region of hBRM/hSNF2α significantly suppressed the anchorage-independent growth of SYT-SSX1-expressing 3Y1 cells. To analyze the transcriptional regulation by SYT-SSX1, we established conditional expression system of SYT-SSX1 and examined the gene expression profiles. The down-regulation of potential tumor suppressor DCC was observed among 1,176 genes analyzed by microarray analysis, and semi-quantitative reverse transcription-PCR confirmed this finding. These data clearly demonstrate transforming activity of human oncogene SYT-SSX1 and also involvement of chromatin remodeling factor hBRM/hSNF2α in human cancer.",
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AU - Nagai, Makoto

AU - Tanaka, Shinya

AU - Tsuda, Masumi

AU - Endo, Shuichi

AU - Kato, Hiroyuki

AU - Sonobe, Hiroshi

AU - Minami, Akio

AU - Hiraga, Hiroaki

AU - Nishihara, Hiroshi

AU - Sawa, Hirofumi

AU - Nagashima, Kazuo

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