RETINOID RECEPTOR CONTROL IN CYTODIFFERENTIATION

拟议研究的一个初步目标是阐明 维甲酸结合蛋白(RBPs)的完整基因组序列， 细胞视黄醇结合蛋白(CRBP) (CRBP)、细胞维甲酸结合蛋白(CRABP)和 膜相关维甲酸结合蛋白(MRABP)。限制性商业惯例将是 纯化至均一，并对其氨基末端进行测序。氨基 这些蛋白质的酸序列和来自一种 已知的高度同源的蛋白质将是合成 几种寡核苷酸。这些寡核苷酸将被用作主要的 在筛选完整的小鼠cDNA文库中的探针；衍生的cDNA将 用于鉴定cRBP、cRABP和mRABP的完整基因组克隆。 逆转录病毒将被构建为包含正常或倒置的 (反义)基因组序列(来自或靠近氨基末端 延伸3个素数以包括所解析的RBP的任何主要内含子 基因。这些逆转录病毒将被感染到已经被 被选为(I)它们的限制性商业惯例cRABP+、cRBP或cRABP+/cRBP+的表达 (Ii)它们对相对低浓度的维甲酸的反应能力 视黄酸+，或视黄醇+。细胞特性的变化(例如， 增殖、转化、终末分化) 和缺少(I)额外表达的RBP基因组序列或 反义信息；(Ii)维甲酸，和(Iii)肿瘤促进剂， 还将对12-O-十四烷基佛波醇-13-乙酸酯进行评价。这些 实验将(I)提供完整的氨基酸和基因组 CRBP、cRABP和MRABP的序列；(Ii)确定限制性商业惯例的要求 在细胞分化的控制中，(Iii)允许RBP的特征 次级构造；及(Iv)为未来发展提供数据 抗维A酸类化合物和抗RBP化合物可能在 化学预防。

An initial objective of the proposed research is the elucidation of the complete genomic sequences of the retinoid-binding proteins (RBPs), cellular retinol-binding protein (cRBP), cellular retinol-binding protein (cRBP), cellular retinoic acid-binding protein (cRABP) and membrane-associated retinoic acid-binding protein (mRABP). RBPs will be purified to homogeneity and their amino-terminal ends sequenced. The amino acid sequences of these proteins and a codon frequency derived from a known, highly homologous protein will be the basis for the synthesis of several oligonucleotides. These oligonucleotides will be used as primary probes in screening complete mouse cDNA libraries; the derived cDNAs will be used to identify the complete genomic clone of cRBP, cRABP, and mRABP. Retroviruses will be constructed that contain either a normal or inverted (anti-sense) genomic sequence (from or near the amino terminal end extending 3 prime to include any primary intron) of the resolved RBP genes. These retroviruses will be infected into cell lines that have been selected for (i) their expression of RBPs cRABP+, cRBP or cRABP+/cRBP+ and (ii) their ability to respond to relatively low concentrations of retinoids retinoic acid+, or retinol+. Changes in cell characteristics (e.g., proliferation, transformation, terminal differentiation) in the presence and absence of (i) an additionally expressed RBP genomic sequence or anti-sense message; (ii) retinoids, and (iii) the tumor promoter, 12-O-tetradecanoylphorbol-13-acetate, will also be evaluated. These experiments will (i) provide the complete amono acid as well as genomic sequence for cRBP, cRABP and MRABP; (ii) establish the requirement for RBPs in the control of cytodifferentiation, (iii) allow characterization of RBP secondary structures; and (iv) provide data for the prospective development of anti-retinoid and anti-RBP compounds that may be of significant value in chemoprevention.