Swiston J, Hubberstey A, Yu G, Young D
Department of Medical Biochemistry, University of Calgary Health Science Centre, Alberta, Canada.
Gene. 1995 Nov 20;165(2):273-7. doi: 10.1016/0378-1119(95)00522-8.
We previously reported the identification of the human CAP and CAP2 genes which encode proteins related to the yeast adenylyl cyclase (CYR)-associated CAP protein. The rat CAP homolog, MCH1, has also been previously cloned. We have cloned a cDNA encoding the rat homolog of CAP2. Rat CAP/MCH1 and CAP2 are 63% identical to each other. Using the reverse transcription-polymerase chain reaction (RT-PCR) method, we have examined CAP/MCH1 and CAP2 mRNA levels in various adult rat tissues. Our results show a dramatic difference in the pattern of expression of these two genes. Consistent with previous reports, we detected CAP/MCH1 mRNA in all tissues examined; however, levels vary substantially between tissues. In particular, we found that CAP/MCH1 mRNA are present at relatively high levels in spleen, testes and lung, at moderate levels in brain, kidney, liver and small intestine, and at significantly lower levels in heart, skeletal muscle and skin. We have also investigated the levels of CAP/MCH1 in rat tissues by immunoblotting with a polyclonal antibody raised against a human CAP::GST fusion protein. In general, we find that the CAP/MCH1 mRNA levels reflect the amount of CAP/MCH1 found in different tissues. In contrast, CAP2 transcripts were present at relatively high levels in testes, at moderate levels in brain, heart and skeletal muscle, at lower levels in lung, skin, kidney and small intestine, and were undetectable in liver or spleen. The differences between the sequences and expression patterns of CAP/MCH1 and CAP2 are significant and suggest that these proteins have distinct functional roles.
我们先前报道了人类CAP和CAP2基因的鉴定,这两个基因编码的蛋白质与酵母腺苷酸环化酶(CYR)相关的CAP蛋白有关。大鼠CAP的同源物MCH1也已于先前被克隆。我们克隆了一个编码大鼠CAP2同源物的cDNA。大鼠CAP/MCH1和CAP2彼此间有63%的同源性。利用逆转录-聚合酶链反应(RT-PCR)方法,我们检测了成年大鼠各种组织中CAP/MCH1和CAP2的mRNA水平。我们的结果显示这两个基因的表达模式存在显著差异。与先前的报道一致,我们在所有检测的组织中都检测到了CAP/MCH1 mRNA;然而,不同组织间的水平差异很大。特别是,我们发现CAP/MCH1 mRNA在脾脏、睾丸和肺中含量相对较高,在脑、肾、肝和小肠中含量中等,而在心脏、骨骼肌和皮肤中含量显著较低。我们还通过用针对人CAP::GST融合蛋白产生的多克隆抗体进行免疫印迹,研究了大鼠组织中CAP/MCH1的水平。总体而言,我们发现CAP/MCH1 mRNA水平反映了不同组织中CAP/MCH1的含量。相比之下,CAP2转录本在睾丸中含量相对较高,在脑、心脏和骨骼肌中含量中等,在肺、皮肤、肾和小肠中含量较低,在肝脏或脾脏中未检测到。CAP/MCH1和CAP2的序列及表达模式差异显著,表明这些蛋白质具有不同的功能作用。