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首页> 外文期刊>Molecular and Cellular Biology >Transcriptional stimulation of the retina-specific QR1 gene upon growth arrest involves a Maf-related protein.
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Transcriptional stimulation of the retina-specific QR1 gene upon growth arrest involves a Maf-related protein.

机译：生长停滞时视网膜特异性QR1基因的转录刺激涉及Maf相关蛋白。

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The avian neural retina (NR) is derived from proliferating neuroectodermal precursors which differentiate after terminal mitosis and become organized in cell strata. Proliferation of postmitotic NR cells can be induced by infection with Rous sarcoma virus (RSV) and requires the expression of a functional v-Src protein. QR1 is a retina-specific gene expressed exclusively at the stage of growth arrest and differentiation during retinal development. In NR cells infected with tsPA101, an RSV mutant conditionally defective in pp60v-src mitogenic capacity, QR1 expression is downregulated in proliferating cells at 37 degrees C and is fully restored when the cells become quiescent as a result of pp60v-src inactivation at 41 degrees C. We were able to arrest proliferation of tsPA101-infected quail NR cells expressing an active v-Src protein by serum starvation at 37 degrees C. This allowed us to investigate the role of cell growth in regulating QR1 transcription. We report that QR1 transcription is stimulated in growth-arrested cells at 37 degrees C compared with that in proliferating cells maintained at the same temperature. Growth arrest-dependent stimulation of QR1 transcription requires the integrity of the A box, a previously characterized cis-acting element responsible for QR1 transcriptional stimulation upon v-Src inactivation and during retinal differentiation. We also show that formation of the C1 complex on the A box is increased upon growth arrest by serum starvation in the presence of an active v-Src oncoprotein. Thus, the C1 complex represents an important link between cell cycle and developmental control of QR1 gene transcription during NR differentiation and RSV infection. By using antibodies directed against different Maf proteins of the leucine zipper family and competition with Maf consensus site-containing oligonucleotides in a gel shift assay, we show that the C1 complex is likely to contain a Maf-related protein. We also show that a purified bacterially expressed v-Maf protein is able to bind the A box and that the level of a 43-kDa Maf-related protein is increased upon growth arrest in infected retinal cells. Moreover, ectopic expression of c-mafI, c-mafII, and mafB cDNAs in quiescent tsPA101-infected quail NR cells is able to stimulate transcription of a QR1 reporter gene through the A box. Therefore, QR1 appears to be the first target gene for a Maf-related protein(s) in the NR.
机译：鸟类神经视网膜（NR）来源于增殖的神经外胚层前体，它们在末端有丝分裂后分化并在细胞层中组织化。劳斯肉瘤病毒（RSV）感染可诱导有丝分裂后NR细胞增殖，并需要表达功能性v-Src蛋白。 QR1是一种视网膜特异性基因，仅在视网膜发育过程中的生长停滞和分化阶段表达。在感染了tsPA101（一种在pp60v-src有丝分裂能力中有条件缺陷的RSV突变体）的NR细胞中，QR1表达在37℃的增殖细胞中被下调，并且当由于pp60v-src在41度的失活而导致细胞静止时，QR1的表达被完全恢复。 C.我们能够通过在37摄氏度的血清饥饿来阻止表达活性v-Src蛋白的tsPA101感染的鹌鹑NR细胞的增殖。这使我们能够研究细胞生长在调节QR1转录中的作用。我们报告说，与维持在相同温度下的增殖细胞相比，QR1转录在37℃的生长停滞细胞中受到刺激。 QR1转录的生长停滞依赖性刺激需要A盒的完整性，A盒是先前表征的顺式作用元件，负责v-Src失活和视网膜分化期间的QR1转录刺激。我们还显示，在活性v-Src癌蛋白存在下，血清饥饿导致生长停滞，A盒上C1复合物的形成增加。因此，C1复合体代表细胞周期与NR分化和RSV感染过程中QR1基因转录的发育控制之间的重要联系。通过使用针对亮氨酸拉链家族的不同Maf蛋白的抗体，并在凝胶移位试验中与包含Maf共有位点的寡核苷酸竞争，我们显示C1复合物可能包含与Maf相关的蛋白。我们还表明，纯化的细菌表达的v-Maf蛋白能够结合A盒，并且在感染的视网膜细胞中生长停滞后，与43 kDa Maf相关的蛋白的水平会增加。此外，c-mafI，c-mafII和mafB cDNA在静止的tsPA101感染的鹌鹑NR细胞中的异位表达能够通过A盒刺激QR1报告基因的转录。因此，QR1似乎是NR中Maf相关蛋白的第一个靶基因。

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