Hepatoma-derived growth factor represses SET and MYND domain containing 1 gene expression through interaction with C-terminal binding protein.
Hepatoma-derived growth factor represses SET and MYND domain containing 1 gene expression through interaction with C-terminal binding protein.
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DOI：
10.1016/j.jmb.2008.12.080
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发表时间：
2009-03-06
影响因子：
5.6
通讯作者：
Everett, Allen D.
中科院分区：
文献类型：
--
作者：
Yang, Jun;Everett, Allen D.
肝癌衍生生长因子（HDGF）是一种具有促有丝分裂和血管生成活性的核蛋白，在发育中的心脏和脉管系统中高度表达。迄今为止，HDGF 的功能机制尚不清楚。寡核苷酸微阵列分析用于深入了解 HDGF 功能。 HDGF 的腺病毒表达显着（≥ 2 倍）下调一大组（66）个基因，并增加相对少数基因（9）的表达。通过实时PCR验证了两组涉及心血管发育和转录调控的靶基因，包括骨骼/心肌特异性SET和MYND结构域含有1（SMYD1）基因。这表明 HDGF 可以起到转录抑制因子的作用。在单杂交系统中，GBD-HDGF 以剂量依赖性方式显着抑制报告基因活性。这证明HDGF具有转录抑制活性。此外，在 G-7 成肌细胞中，GFP-HDGF 融合体的过度表达特异性下调 SMYD1 mRNA 表达和人 SMYD1 启动子的活性。 HDGF 通过与转录辅阻遏物 C 端结合蛋白 (CtBP) 相互作用来抑制 SMYD1 基因转录。 CtBP 的过表达增强了 HDGF 的反式抑制活性；另一方面，敲除CtBP会减弱HDGF的反式抑制作用。 HDGF 通过 PWWP 结构域内的非规范结合基序 (PKDLF) 结合 CtBP，因为 DL 到 AS 的替代突变消除了 HDGF 和 CtBP 相互作用，并减弱了 HDGF 的反式抑制作用，而不影响 DNA 结合。最后，荧光显微镜研究表明，HDGF 诱导 CtBP 的核积累，表明 HDGF 与 CtBP 形成转录复合物。综上所述，我们的数据表明 HDGF 通过与转录辅阻遏物 CtBP 相互作用，充当 SMYD1 基因的转录阻遏物。由于 HDGF 家族成员中 CtBP 结合基序的适度保守，CtBP 介导的反式抑制活性可能是 HDGF 蛋白的常见功能。
Hepatoma Derived Growth Factor (HDGF) is a nuclear protein with both mitogenic and angiogenic activity, it is highly expressed in the developing heart and vasculature. To date the mechanisms of HDGF’s function are unknown. Oligonucleotide microarray analysis was used to gain insights into HDGF function. Adenoviral expression of HDGF significantly (≥ 2 fold) downregulated a large group (66) of genes, and increased expression of a relatively small number of genes (9). Two groups of target genes which are involved in cardiovascular development and transcriptional regulation were validated by real time PCR, including the skeletal/cardiac muscle specific SET and MYND domain containing 1 (SMYD1) gene. This suggested that HDGF could function as a transcriptional repressor. In a one-hybrid system, GBD-HDGF significantly repressed reporter gene activity in a dose dependent manner. This demonstrated that HDGF has transcriptional repressive activity. Moreover, in G-7 myoblast cells, overexpression of a GFP-HDGF fusion specifically downregulated SMYD1 mRNA expression and the activity of the human SMYD1 promoter. HDGF repressed SMYD1 gene transcription through interaction with a transcriptional corepressor C-terminal binding protein (CtBP). Overexpressing of CtBP potentiated the trans-repressive activity of HDGF; on the other hand, knocking down CtBP attenuated the trans-repressive effect of HDGF. HDGF binds CtBP through a non-canonical binding motif (PKDLF) within the PWWP domain, as substitutional mutation of DL to AS abolished HDGF and CtBP interaction and diminished the trans-repressive effect of HDGF without affecting DNA binding. Finally, fluorescent microscopy studies showed that HDGF induced the nuclear accumulation of CtBP suggesting that HDGF forms a transcriptional complex with CtBP. Taken together, our data demonstrate that HDGF functions as a transcriptional repressor of the SMYD1 gene, through interaction with the transcriptional corepressor CtBP. Because of moderate conservation of the CtBP binding motif in HDGF family members, trans-repressive activity mediated by CtBP may be a common function among HDGF proteins.
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影响因子：
4.1
作者：
Dietz, F;Franken, S;Gieselmann, V
通讯作者：
Gieselmann, V
影响因子：
2.5
作者：
Everett, AD
通讯作者：
Everett, AD
影响因子：
8
作者：
Nameki, N;Tochio, N;Yokoyama, S
通讯作者：
Yokoyama, S
DOI：
10.1006/bbrc.1999.1733
发表时间：
1999-12-09
影响因子：
3.1
作者：
Ikegame, K;Yamamoto, M;Nakamura, H
通讯作者：
Nakamura, H
影响因子：
4.8
作者：
Bachman, KE;Rountree, MR;Baylin, SB
通讯作者：
Baylin, SB