Leyva Leyva M, Gariglio P, Rangel L M, Valdés J, Ayala P, Orozco E
Department of Experimental Pathology, CINVESTAV-IPN, México, D.F.
Parasitol Res. 1993;79(2):153-9. doi: 10.1007/BF00932262.
The evolutionarily conserved proto-oncogene c-myc is involved in both proliferation and differentiation processes of higher eukaryotic cells. We report here the identification and characterization of sequences homologous to c-myc in different Entamoeba species using a fragment of the mammalian c-myc gene as a probe. This probe hybridized with fragments of 3.5 and 3.4 kilobases (kb) in E. histolytica HindIII of EcoRI digested DNA. In E. invadens it recognized fragments of 3.1 and 2.8 kb, and in Laredo strain (reported as E. moshkovskii by Clark and Diamond in 1991) the probe hybridized with fragments of 17 kb. The c-myc probe identified transcripts of 3.3 and 1.5 kb in E. histolytica, transcripts of 1.8 and 1.3 kb in Laredo strain, and transcripts of 3.7, 1.8, 1.5 and 1.1 kb in E. invadens. Antibodies against a highly conserved region of the c-myc protein recognized in E. histolytica polypeptides of 35, 40, and 60 kDa. The expression of the 60 kDa polypeptide was temperature-inducible in Laredo strain. In E. invadens a 110 kDa strong band was detected by the antibodies. Surprisingly, E. invadens myc-like sequences and proteins showed greater homology to mammalian c-myc gene and proteins. Expression of proteins antigenically related to c-myc varied according to the cell cycle phase of E. histolytica. These proteins peaked during D, G1, and S phases and declined during G2.
进化上保守的原癌基因c-myc参与高等真核细胞的增殖和分化过程。我们在此报告,使用哺乳动物c-myc基因的片段作为探针,在不同的溶组织内阿米巴物种中鉴定和表征与c-myc同源的序列。该探针与经EcoRI消化的溶组织内阿米巴HindIII DNA中的3.5和3.4千碱基(kb)片段杂交。在侵袭内阿米巴中,它识别出3.1和2.8 kb的片段,而在拉雷多菌株(克拉克和戴蒙德在1991年报告为莫斯科维茨内阿米巴)中,该探针与17 kb的片段杂交。c-myc探针在溶组织内阿米巴中鉴定出3.3和1.5 kb的转录本,在拉雷多菌株中鉴定出1.8和1.3 kb的转录本,在侵袭内阿米巴中鉴定出3.7、1.8、1.5和1.1 kb的转录本。针对c-myc蛋白高度保守区域的抗体识别出溶组织内阿米巴中35、40和60 kDa的多肽。60 kDa多肽的表达在拉雷多菌株中受温度诱导。在侵袭内阿米巴中,抗体检测到一条110 kDa的强条带。令人惊讶的是,侵袭内阿米巴的类myc序列和蛋白与哺乳动物c-myc基因和蛋白显示出更高的同源性。与c-myc抗原相关的蛋白表达根据溶组织内阿米巴的细胞周期阶段而变化。这些蛋白在D、G1和S期达到峰值,在G2期下降。