Immunoselection of cDNAs to avian intestinal calcium binding protein 28K and a novel calmodulin-like protein: assessment of mRNA regulation by the vitamin D hormone.
Immunoselection of cDNAs to avian intestinal calcium binding protein 28K and a novel calmodulin-like protein: assessment of mRNA regulation by the vitamin D hormone.
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DOI：
10.1021/bi00399a046
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发表时间：
1987-12
期刊：
影响因子：
2.9
通讯作者：
D. Mangelsdorf;B. Komm;D. McDonnell;J. Pike;M. Haussler
中科院分区：
文献类型：
--
作者：
D. Mangelsdorf;B. Komm;D. McDonnell;J. Pike;M. Haussler
钙在各种细胞过程中的作用已被充分证明。钙的储存、分布和递送由一个结合蛋白家族调节，包括肌钙蛋白C、钙调蛋白、小清蛋白和维生素D依赖性钙结合蛋白（CaBP-28），所有这些都是从共同的祖先基因进化而来的。为了评估维生素D对基因转录的调节，利用多价CaBP-28抗体作为探针，从鸡肠道λ gt 11文库中分离CaBP-28 cDNA（767个碱基对）。与CaBP-28 cDNA的鉴定一致，还分离出一组cDNA（用抗CaBP-28抗体），其与鸡脑钙调素（CaM）具有84%的核苷酸同源性和99%的推导氨基酸同源性。这种新的钙调素样cDNA被命名为neoCaM。CaBP-28 cDNA与neoCaM之间几乎没有核苷酸同源性。CaBP-28 cDNA与2000、2900和3300个碱基的三种转录物杂交，这三种转录物由1，25-二羟基维生素D3 [1，25（OH）2D 3]显著诱导，而neoCaM cDNA识别三种不同的（来自CaBP-28）转录物。这些mRNA中的两个是1400和1800个碱基，如脑钙调素所述，但另一个更大的4000个碱基的转录物与新钙调素检测。无论是钙调素还是新钙调素的转录显示任何调制1，25（OH）2D 3。在这里，我们讨论了可能的意义，不仅是一个单一的抗体，但也有关系的新钙调素的其他良好的特点钙调素的cDNA的cDNA的分离。
Calcium's role in a variety of cellular processes has been well documented. The storage, distribution, and delivery of calcium are regulated by a family of binding proteins including troponin C, calmodulin, parvalbumin, and vitamin D dependent calcium binding protein (CaBP-28), all of which have evolved from a common ancestral gene. To evaluate vitamin D regulation of gene transcription, a CaBP-28 cDNA (767 base pairs) was isolated from a chicken intestine lambda gt11 library utilizing a polyvalent CaBP-28 antibody as a probe. Coincident with the identification of the CaBP-28 cDNA, a group of cDNAs also was isolated (with the anti-CaBP-28 antibody) that demonstrated 84% nucleotide homology and 99% deduced amino acid homology with chicken brain calmodulin (CaM). This new CaM-like cDNA was named neoCaM. There is little nucleotide homology between the CaBP-28 cDNA and neoCaM. The CaBP-28 cDNA hybridizes with three transcripts of 2000, 2900, and 3300 bases which are dramatically induced by 1,25-dihydroxyvitamin D3 [1,25(OH)2D3], while the neoCaM cDNA recognizes three distinct (from CaBP-28) transcripts. Two of these mRNAs are 1400 and 1800 bases as described for brain CaM, but another larger 4000-base transcript is detected with neoCaM. Neither the CaM nor the neoCaM transcript reveals any modulation by 1,25(OH)2D3. Herein, we discuss the possible significance of not only the isolation of both cDNAs with a single antibody but also the relation of neoCaM to other well-characterized CaM cDNAs.