The NF-Y complex negatively regulates Caenorhabditis elegans tbx-2 expression.
The NF-Y complex negatively regulates Caenorhabditis elegans tbx-2 expression.
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DOI：
10.1016/j.ydbio.2013.08.001
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发表时间：
2013-10-01
影响因子：
2.7
通讯作者：
Okkema, Peter G.
中科院分区：
文献类型：
--
作者：
Milton, Angenee C.;Packard, Adelaide V.;Clary, Lynn;Okkema, Peter G.
T-box基因在动物发育过程中经常以动态模式表达，但控制这些基因表达的机制尚不清楚。梭线虫T-box基因tbx-2对于ABA衍生的咽肌的发育、HSN/PHB谱系中神经细胞命运的特化以及嗅觉神经元的适应是必需的。tbx-2的表达模式是复杂的，并且已经描述了在胚胎发育期间在咽前体和体壁肌肉中的表达，以及在成体中在两栖动物感觉神经元和咽神经元中的表达。为了研究调节tbx-2基因表达的机制，我们在含有Ptbx-2：：gfp报告基因的菌株中进行了转录因子基因的RNAi筛选，并鉴定了核因子Y（NF-Y）复合物作为tbx-2表达的负调节因子。NF-Y是由A、B和C亚基组成的异源三聚体CCAAT结合复合物，并且通过RNAi或使用突变体减少NF-Y亚基nfya-1、nfy B-1或nfyc-1导致皮下接缝细胞和肠中的异位Ptbx-2：：gfp表达。tbx-2启动子中两个CCAAT盒的突变导致类似的异位Ptbx-2：：gfp表达模式，表明NF-Y直接抑制tbx-2启动子。tbx-2 mRNA在nfya-1无效突变体中适度增加，表明NF-Y抑制内源性tbx-2的表达。最后，我们确定并表征了第二个位点的突变，提高致死性的温度敏感tbx-2突变体，并表明，这种突变是一个缺失的nfyb-1基因。总之，这些结果确定NF-Y作为体内tbx-2功能的重要调节剂。
T-box genes are frequently expressed in dynamic patterns during animal development, but the mechanisms controlling expression of these genes are not well understood. The C. elegans T-box gene tbx-2 is essential for development of the ABa-derived pharyngeal muscles, specification of neural cell fate in the HSN/PHB lineage, and adaptation in olfactory neurons. The tbx-2 expression pattern is complex, and expression has been described in pharyngeal precursors and body wall muscles during embryogenesis, and amphid sensory neurons and pharyngeal neurons in adults. To examine mechanisms regulating tbx-2 gene expression, we performed an RNAi screen of transcription factor genes in strains containing a Ptbx-2::gfp reporter and identified the Nuclear Factor Y (NF-Y) complex as a negative regulator of tbx-2 expression. NF-Y is a heterotrimeric CCAAT-binding complex consisting of A, B and C subunits, and reduction of the NF-Y subunits nfya-1, nfyb-1, or nfyc-1 by RNAi or using mutants results in ectopic Ptbx-2::gfp expression in hypodermal seam cells and gut. Mutation of two CCAAT-boxes in the tbx-2 promoter results in a similar pattern of ectopic Ptbx-2::gfp expression, suggesting NF-Y directly represses the tbx-2 promoter. tbx-2 mRNA is moderately increased in nfya-1 null mutants, indicating NF-Y represses expression of endogenous tbx-2. Finally we identify and characterize a second-site mutation that enhances lethality of a temperature sensitive tbx-2 mutant and show that this mutation is a deletion in the nfyb-1 gene. Together, these results identify NF-Y as an important regulator of tbx-2 function in vivo.
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影响因子：
11.4
作者：
Martin, Nadine;Benhamed, Moussa;Bischof, Oliver
通讯作者：
Bischof, Oliver
影响因子：
2.7
作者：
Chowdhuri, Sinchita Roy;Crum, Tanya;Okkema, Peter G.
通讯作者：
Okkema, Peter G.
影响因子：
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作者：
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通讯作者：
Salzberg, Steven L.
DOI：
10.1016/j.bbcan.2010.07.001
发表时间：
2010-12
期刊：
Biochimica et biophysica acta
影响因子：
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作者：
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通讯作者：
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影响因子：
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作者：
Buschlen, S;Amillet, JM;Bolotin-Fukuhara, M
通讯作者：
Bolotin-Fukuhara, M