A Control Mechanism of Gene and Protein Expression in Normal and Abnormal Red Cell Membranes

正常和异常红细胞膜基因和蛋白质表达的控制机制

• 批准号: 09044346
• 负责人: YAWATA Yoshihito
• 金额: $ 4.61万
• 依托单位: KAWASAKI MEDICAL SCHOOL
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for international Scientific Research
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09044346/
• 关键词: Gene expression; Gene methylation; CpG site; Hereditary spherocytosis; Allele 4.1 (-) Madrid; Molecular electron microscopy; Gene analysis; Cytoskeletal network; allele 4.1(-)Madrid

(I) The state of methylation on the genes of red cell membrane proteins :The state of methylation was studied at the promoter region of the genes assigned to human red cell membrane proteins (beta-spectrin, band 3, and protein 4.2) by the bisulfite method (Zeschnigk M et al.1997). In normal condition, the 13 sites of 5'-CpG-3' on the genes of band 3 (B3) and the 7 CpG sites of protein 4.2 (P4.2) were heavily methylated in the peripheral mononuclear cells. In contrast, the 40 sites of the beta-spectrin gene were totally unmethylated. In addition, in total deficiency of P4.2, the 4 CpG sites of the B3 gene were specifically unmethylated (Yawata Y,et al : Blood 90 : 8b, 1997 ; Wada H,et al : Exp Hematol, in press, 1998).(II) Phenotypic abnormalities of human red cell membrane disorders :Red cell membrane proteins and their related genes were analyzed in 92 cases of the 37 kindreds of hereditary spherocytosis and in 36 cases of normal subjects. In these patients, 3 frameshift mutations, 5 missense mutations, and 7 polymorphism were identified on the B3 gene, contrary to 12 silent mutations on the ankyrin gene (Kanzaki A,et al : Brit J Haematol 99 : 522-530,1997 ; Kanzaki A,et al : Blood 90 : 6b-7b, 1997). Markedly deranged cytoskeletal network was detected in the red cell membrane in situ with allele 4.1 (-) Madrid by immuno-electron microscopy (Yawata A,et al : Blood 90 : 2471-2481,1997).

(I)红细胞膜蛋白基因的甲基化状态：用亚硫酸氢法(Zeschnigk M et al.1997)研究了红细胞膜蛋白基因(β-Spectrin，带3和蛋白4.2)启动子区域的甲基化状态。在正常情况下，外周血单核细胞中蛋白4.2基因的13个5‘-CpG-3’位(B3)和7个CpG位(P4.2)发生了高度甲基化。相比之下，β-血影蛋白基因的40个位点完全没有甲基化。此外，在P4.2的完全缺失中，B3基因的4个CpG位点被特异性地去甲基化(Yawata Y等人：血液90：8B，1997；Wada H等：EXP hematol，1998)。(Ii)人类红细胞膜疾病的表型异常：对37个遗传性球形细胞增多症家系中的92例和36例正常人的红细胞膜蛋白及其相关基因进行了分析。在这些患者中，在B3基因上发现了3个移码突变、5个错义突变和7个多态，而在ankyrin基因上发现了12个沉默突变(Kanzaki A等人：Brit J Haematol 99：522-530,1997；Kanzaki A等人：血液90：6B-7b，1997)。免疫电子显微镜检测到红细胞膜上有明显错乱的细胞骨架网络，其等位基因为4.1(-)马德里(Yawata A等：血液90：2471-2481,1997)。

项目成果

八幡 義人: "新臨床内科学第7版" 医学書院,東京, 3 (1997)

八幡义人：《新临床内科第 7 版》Igakushoin，东京，3 (1997)

Wada, H., Kanzaki, A., Yawata, A., Kaku, M., Takezono, M., Inoue, T., Sugihara, T., Yamada, O., Yawata, Y., Uchikawa, M., Fujimoto, T., Fujimura, K.: "A novel combined red cell membrane anomaly of abnormal glycoproteins (GP-A/band 3) and disrupted cytoske

和田 H.、神崎 A.、八幡 A.、加来 M.、竹园 M.、井上 T.、杉原 T.、山田 O.、八幡 Y.、内川 M.、

Yawata A, et al: "A markedly disrupted skeletal network with abnormally distributed intramembrane particles in complete protein 4.1-deficient red blood cells (allele Madrid): Implications regarding a critical role of protein 4.1 in maintenance of the inte

Yawata A 等人：“在完整蛋白 4.1 缺陷的红细胞（马德里等位基因）中，骨骼网络明显破坏，膜内颗粒分布异常：关于蛋白 4.1 在维持骨骼网络中的关键作用的影响

Kanzaki A, et al: "Total absence of protein 4.2 and partial deficiency of band 3 in hereditary spherocytosis"Brit.J.Haematol.. 99. 522-530 (1997)

Kanzaki A 等人：“遗传性球形红细胞增多症中蛋白质 4.2 完全缺失且条带 3 部分缺乏”Brit.J.Haematol.. 99. 522-530 (1997)

Yawata, Y., et al: "Quantitative electron microscopy demonstrates an excellent correlation with the extent of truncation of C-terminal region of human erythroid β-spectrins by exon skipping." Blood. 90(Supple.1). 8b (1997)

Yawata, Y. 等人：“定量电子显微镜显示与外显子跳跃导致的人红系 β-血影蛋白 C 末端区域的截断程度具有良好的相关性。”(Supple.1)。