Quantification and regulation of mRNAs encoding beaded filament proteins in the chick lens
Quantification and regulation of mRNAs encoding beaded filament proteins in the chick lens
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DOI：
10.1076/ceyr.16.8.838.8990
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发表时间：
1997-08-01
影响因子：
2
通讯作者：
Poosch, MS
中科院分区：
文献类型：
--
作者：
Ireland, ME;Goebel, DJ;Poosch, MS
目的。目的：定量鸡晶状体区域珠丝蛋白mRNA的表达水平，并利用细胞培养技术研究其在体外对信息和蛋白水平的调控。采用RNase保护实验和Northern印迹法定量解剖晶状体中珠状细丝蛋白mRNA水平。培养细胞经cAMP类似物处理后，采用RNase保护法检测mRNA含量，采用Western blotting和ELISA技术检测蛋白质含量。与环状垫细胞相比，皮质纤维细胞中的珠状丝蛋白信息水平显著上调。在核纤维细胞中也检测到全长信息。存在一种不寻常形式的CP49信息，带有层粘连状插入。CP49（INS）也成立了。细胞内cAMP水平升高，信息和蛋白水平均受调控。纤维细胞发育过程中珠状丝蛋白水平的积累可能与camp介导的信息转录增加有关，CP49（INS）的存在可能为研究中间丝的组装机制提供新的思路。
Purpose. To quantify the expression of beaded filament protein mRNA levels in regions of the chick lens and to examine the in vitro regulation of message and protein levels using cell culture techniques.Methods. RNase protection assays and Northern blotting were used to quantify beaded filament protein mRNA levels in dissected lenses. Cultured cells were assayed for mRNA with RNase protection and for protein with Western blotting and ELISA techniques after treatment with cAMP analogs.Results. Beaded filament protein message levels were greatly up-regulated in cortical fiber cells compared to annular pad cells. Full length messages were also detected in nuclear fiber cells. The presence of an unusual form of the CP49 message with a lamin-like insert. CP49(INS), was also established. Both message and protein levels were subject to regulation in response to elevated intracellular cAMP levels.Conclusions. The accumulation of beaded filament protein levels during fiber cell development may be due to the increased cAMP-mediated transcription of message, The presence of CP49(INS) may lend new insight into mechanisms of intermediate filament assembly.