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首页> 外文期刊>Molecular and Cellular Biology >Differential regulation of trypsinogen mRNA translation: full-length mRNA sequences encoding two oppositely charged trypsinogen isoenzymes in the dog pancreas.
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Differential regulation of trypsinogen mRNA translation: full-length mRNA sequences encoding two oppositely charged trypsinogen isoenzymes in the dog pancreas.

机译：胰蛋白酶原mRNA翻译的差异调节：犬胰脏中编码两个带相反电荷的胰蛋白酶原同工酶的全长mRNA序列。

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摘要

In the absence of changes in functional mRNA levels, stimulation of the pancreas with caerulein, a peptide analog of cholecystokinin, has been previously shown to increase the synthesis of anionic but not cationic trypsinogen. To look for structure-function correlations, a high-yield, full-length cDNA library has been constructed from canine pancreatic poly(A)+ mRNA. Full-length clones coding for the two major trypsinogen isoenzyme forms have been identified by colony hybridization and verified by in vitro translation of hybrid-selected mRNA in the presence of microsomal membranes and an optimal redox potential. Disulfide-bonded translation products were separated and identified by two-dimensional isoelectric focusing-sodium dodecyl sulfate-gel electrophoresis. Nucleotide sequence analysis allowed us to deduce the amino acid sequences for the anionic and cationic forms of canine trypsinogen, which contain 232 and 231 residues, respectively (77% amino acid identity), and the 15-residue amino terminal signal sequences (53% amino acid identity) associated with the two presecretory forms. Measurements of relative and absolute mRNA levels, when related to relative protein synthesis values, indicated that the translational efficiency of anionic trypsinogen mRNA exceeded that of cationic trypsinogen mRNA by 1.5- to 2.9-fold under basal conditions. Analysis of the 5' noncoding regions of trypsinogen mRNAs revealed a striking conservation of sequence (10 of 12 bases) between dog and rat anionic trypsinogen forms. This contrasted markedly with the divergence of the 5' noncoding regions observed between dog anionic and cationic trypsinogen mRNAs.
机译：在功能性mRNA水平没有变化的情况下，以前已经证明用胆囊收缩素的肽类似物caerulein刺激胰腺可以增加阴离子而不是阳离子胰蛋白酶原的合成。为了寻找结构功能的相关性，已经从犬胰腺poly（A）+ mRNA构建了高产量，全长的cDNA文库。已经通过菌落杂交鉴定了编码两种主要胰蛋白酶原同工酶形式的全长克隆，并通过在微粒体膜和最佳氧化还原电势存在下杂交选择的mRNA的体外翻译进行了验证。分离并通过二维等电聚焦-十二烷基硫酸钠-凝胶电泳鉴定二硫键翻译产物。核苷酸序列分析使我们能够推断出犬胰蛋白酶原的阴离子和阳离子形式的氨基酸序列，分别包含232和231个残基（77％的氨基酸同一性）和15个残基的氨基末端信号序列（53％的氨基酸）酸身份）与这两种分泌形式有关。相对和绝对mRNA水平的测量，当与相对蛋白质合成值相关时，表明在碱性条件下，阴离子胰蛋白酶原mRNA的翻译效率比阳离子胰蛋白酶原mRNA的翻译效率高1.5到2.9倍。胰蛋白酶原mRNA的5'非编码区的分析表明，在狗和大鼠阴离子胰蛋白酶原形式之间，序列具有惊人的保守性（12个碱基中的10个碱基）。这与在狗阴离子和阳离子胰蛋白酶原mRNA之间观察到的5'非编码区的差异形成鲜明对比。

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