Kwon Yunjeong, Ko Jeong Heon, Byung-Gyu Kim, Kim Myoung Hee
Department of Anatomy and Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine, C.P.O. Box 8044, Seoul, 120-752, Korea.
Mol Biol Rep. 2003 Sep;30(3):141-8. doi: 10.1023/a:1024920418148.
Although Hox genes are known to mediate developmental decisions involved in pattern formation during embryogenesis, it is still not well understood what Hox regulates. In order to analyze Hoxc8 downstream target genes, a stable cell line overexpressing Hoxc8 was established using F9 murine teratocarcinoma cells, proteom samples were analyzed by 2-DE, and compared with controls. The protein spots having differences more than 4 fold in intensity were selected, analyzed by MALDI-TOF, and grouped in terms of putative function; cytoskeleton and motility (vimentin, gamma-actin, tropomyosin, and tubulin beta-5 chain); folding, modification and degradation of protein (GRP78, proteasome subunit alpha type 5, 26S proteasome regulatory subunit p27 protein, and PDIR); metabolism (ATP synthase beta subunit, Pgam1, and CAII); transcription/translation factors and general nucleic acid binding proteins (RbAp46, PCNA, eEF-1-beta, and nucleophosmin). Although it may not be significant, 50% of the genes were located on chromosomes 2 and 3, suggesting the possibility of a non-random distribution of Hox downstream genes. Almost 50% of the genes analyzed showed some relation with Hox protein directly or indirectly; i.e., tubulin beta 5, EF-1 beta and PCNA have been reported to contain putative Hox binding regulatory sites and genes like vimentin, pgam1 and nucleophosmin to be regulated by RA, a potent modulator of Hox expression. These results altogether imply that proteom analysis could be a possible tool for the analysis of the potent Hox realizator genes, which provides a new insight into the function of Hox on pattern formation during embryogenesis.
尽管已知Hox基因在胚胎发生过程中参与模式形成的发育决策介导，但Hox调控的具体机制仍未完全明确。为了分析Hoxc8的下游靶基因，利用F9小鼠畸胎瘤细胞建立了过表达Hoxc8的稳定细胞系，通过二维电泳分析蛋白质组样本，并与对照进行比较。选择强度差异超过4倍的蛋白质斑点，用基质辅助激光解吸电离飞行时间质谱进行分析，并根据推定功能进行分组；细胞骨架与运动（波形蛋白、γ-肌动蛋白、原肌球蛋白和微管蛋白β-5链）；蛋白质的折叠、修饰和降解（葡萄糖调节蛋白78、蛋白酶体α5亚基、26S蛋白酶体调节亚基p27蛋白和蛋白质二硫键异构酶相关蛋白）；代谢（ATP合酶β亚基、磷酸甘油酸变位酶1和碳酸酐酶II）；转录/翻译因子和一般核酸结合蛋白（视网膜母细胞瘤结合蛋白46、增殖细胞核抗原、真核生物延伸因子1-β和核磷蛋白）。尽管这可能并不显著，但50%的基因位于2号和3号染色体上，这表明Hox下游基因存在非随机分布的可能性。几乎50%的分析基因直接或间接与Hox蛋白存在某种关联；例如，据报道微管蛋白β5、延伸因子1-β和增殖细胞核抗原含有推定的Hox结合调控位点，波形蛋白、磷酸甘油酸变位酶1和核磷蛋白等基因受视黄酸调控，视黄酸是Hox表达的有效调节剂。这些结果共同表明，蛋白质组分析可能是分析Hox效应基因的一种可行工具，这为深入了解Hox在胚胎发生过程中模式形成的功能提供了新的视角。
