Kim W K, Baluda M A
Department of Pathology, School of Medicine, University of California, Los Angeles 90024.
Mol Cell Biol. 1989 Sep;9(9):3771-6. doi: 10.1128/mcb.9.9.3771-3776.1989.
Comparison of the nucleotide sequence of the upstream c-myb exon UE3 with the sequences of a thymus c-myb cDNA and of a B-lymphoma c-myb cDNA suggested the existence of T- and B-cell-specific heterogeneity in the 5'-terminal region of the c-myb coding sequence. This possibility was investigated with T-cell-specific and B-cell-specific DNA probes in a Northern (RNA) blot analysis of mRNAs from different hematopoietic cell types and from chicken embryo fibroblasts. The hematopoietic tissues analyzed were bone marrow, bursa of Fabricius, and thymus from 1-day-old chicks, 13-day yolk sac, and spleen from 16-day embryos. At least three different c-myb mRNA species were found to have 5'-terminal heterogeneity that was specific for either B cells, T cells, or the other hematopoietic cells and chicken embryo fibroblasts. This lineage-specific heterogeneity in the c-myb transcript was found to be expressed in the bone marrow precursors of B and T cells before they migrated to their definitive differentiation sites. S1 nuclease protection analysis of the UE3 exon, part of which appeared to be coding sequences for thymic c-myb mRNA, revealed that this exon is utilized either in its entirety or partially in a cell-lineage-specific manner by all six tissues analyzed. Also, the 5'-terminal exon(s) present in the thymus cDNA was absent in c-myb mRNAs from the other cell types analyzed.
将上游c-myb外显子UE3的核苷酸序列与胸腺c-myb cDNA及B淋巴瘤c-myb cDNA的序列进行比较,结果表明在c-myb编码序列的5'末端区域存在T细胞和B细胞特异性的异质性。利用T细胞特异性和B细胞特异性DNA探针,对来自不同造血细胞类型及鸡胚成纤维细胞的mRNA进行Northern(RNA)印迹分析,对这一可能性进行了研究。所分析的造血组织包括1日龄雏鸡的骨髓、法氏囊和胸腺、13日龄卵黄囊以及16日龄胚胎的脾脏。结果发现,至少有三种不同的c-myb mRNA存在5'末端异质性,这种异质性分别特异于B细胞、T细胞或其他造血细胞及鸡胚成纤维细胞。c-myb转录本中的这种谱系特异性异质性在B细胞和T细胞迁移至其最终分化位点之前的骨髓前体细胞中即已表达。对UE3外显子进行S1核酸酶保护分析,结果显示该外显子的一部分似乎是胸腺c-myb mRNA的编码序列,在所分析的所有六种组织中,该外显子全部或部分以细胞谱系特异性方式被利用。此外,胸腺cDNA中存在的5'末端外显子在所分析的其他细胞类型的c-myb mRNA中并不存在。