GENE REGULATION OF RAT DENTIN SIALOPROTEIN

大鼠牙本质唾液酸蛋白的基因调控

• 批准号: 2458644
• 负责人: HELENA H Ritchie
• 金额: $ 0.01万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-09-30 至 1999-01-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2458644
• 关键词: DNA footprinting; complementary DNA; dentin; dentinogenesis; developmental genetics; gene induction /repression; genetic library; genetic promoter element; in situ hybridization; laboratory rat; luciferin monooxygenase; northern blottings; odontoblasts; pharmacology; polymerase chain reaction; tissue /cell culture; transcription factor; transfection; transforming growth factors; vitamin D

Although the mechanisms involved in dentinogenesis are unknown, it is clear that a unique set of extracellular matrix (ECM) proteins participates in the formation of predentin and its subsequent mineralization to form dentin. Mature odontoblasts secrete collagen at the cell border and non-collagenous proteins (NCPs) at the mineralization front (possibly through odontoblastic processes). Here NCPs form complexes with collagen. Carbonate apatite crystals are formed and this site-specific process of crystal initiation and growth is believed to be controlled by the collagen-NCP complex. In order to elucidate details of dentinogenesis, additional information relative to the NCPs, their metabolism and gene regulation is needed. Of particular interest are proteins found uniquely in dentin ECM. One dentin specific protein, dentin sialoprotein (DSP), a 53 kDa protein, is a sialic acid-rich protein similar in overall properties to cell attachment proteins bone sialoprotein (BSP) and osteopontin (OPN). However, DSP is synthesized only by odontoblasts and dental pulp and not by osteoblasts or other cell types. The sequence of DSP deduced from its cDNA is dissimilar to those of BSP, OPN and other proteins. DSP contains consensus sequences for N- and O-glycosylation sites, as well as casein kinase and 11 phosphorylation sites. Northern blots detected multiple transcripts of approximately 4.6 kb and 1.5 kb. Recent Southern blot analysis of genomic clones strongly suggests the presence of two related DSP genes. We hypothesize that the synthesis and secretion of DSP is crucial to the formation of healthy dentin. In order to test this hypothesis, we will employ molecular biological techniques to elucidate gene structures and their regulation. We propose the following Specific Aims: i. To determine the genomic organization of DSP gene(s). 2. To characterize multiple DSP transcripts and to examine the spatial and temporal expression of these transcripts. 3. To characterize the promoter(s) of DSP gene(s) including DNA sequence, minimal regulatory sequences and nuclear transcription factors and 4. To study the potential regulation of the DSP gene(s) by signal transducing molecules. The studies outlined in this proposal will provide a solid molecular basis for understanding important regulatory events controlling dentinogenesis.

尽管牙齿发生中涉及的机制尚不清楚，但这是 清楚一组独特的细胞外基质（ECM）蛋白 参加predentin及其后续的形成 矿化形成牙本质。成熟的odontoblasts分泌胶原蛋白 矿化处的细胞边界和非胶原蛋白（NCP） 正面（可能是通过Odontoblastic过程）。这里是NCPS形式 胶原蛋白的复合物。形成了碳酸盐磷灰石晶体，这 据信晶体启动和生长的特定地点的过程是 由胶原-NCP复合物控制。 为了阐明细节 牙齿发生，相对于NCP的其他信息，它们 需要代谢和基因调节。特别感兴趣的是 蛋白质在牙本质ECM中唯一发现。一种牙本质特异性蛋白， 53 kDa蛋白是一种含有唾液酸的乙蛋白唾液蛋白（DSP），是一种富含唾液酸的蛋白 蛋白质在整体特性与细胞附着蛋白骨骨相似 唾液蛋白（BSP）和骨桥蛋白（OPN）。 但是，DSP合成了 仅由Odontoblasts和Dental Pulp，而不是由成骨细胞或其他细胞 类型。从其cDNA推导的DSP的序列与那些 BSP，OPN和其他蛋白质。 DSP包含N-的共识序列 和O-糖基化位点，以及酪蛋白激酶和11 磷酸化位点。北印迹检测到多个成绩单 约4.6 kb和1.5 kb。最近的Southern印迹分析基因组分析 克隆强烈建议存在两个相关的DSP基因。 我们假设DSP的合成和分泌对于 健康牙本质的形成。为了检验这一假设，我们将 采用分子生物学技术来阐明基因结构和 他们的监管。我们提出以下特定目标：i。确定 DSP基因的基因组组织。 2。要表征多个 DSP转录本并检查 这些成绩单。 3。表征DSP基因的启动子 包括DNA序列，最小调节序列和核 转录因子和4。研究DSP的潜在调节 基因通过信号转导分子。其中概述了 提案将为理解重要的分子提供稳定的基础 控制牙本​​质发生的调节事件。