Mano H, Mano K, Tang B, Koehler M, Yi T, Gilbert D J, Jenkins N A, Copeland N G, Ihle J N
Department of Biochemistry, St. Jude Children's Research Hospital, Memphis, Tennessee 38105.
Oncogene. 1993 Feb;8(2):417-24.
The Tec kinase was initially identified as a novel cytoplasmic protein tyrosine kinase that is preferentially expressed in the liver and is highly homologous to the Drosophila Dsrc28C src-related tyrosine kinase. In screening of interleukin 3 (IL-3)-dependent myeloid leukemia cells for protein tyrosine kinases, we observed that all cell lines examined expressed high levels of Tec transcripts. However, characterization of Tec cDNAs indicated that they differed significantly from the published sequence. Most strikingly, an insertion of 41 bp in the 5' region affects the initiation codon and results in replacing the published 13 amino acid amino-terminal sequences with 94 amino acids. Using polymerase chain reaction (PCR) analysis, only the form containing the insertion was detected in hematopoietic cells. In addition, we found an in-frame insertion of 66 bp that introduces an additional 22 amino acids into the SH3 domain. This insertion restores conserved SH3 sequences that are found in the src gene family and in the Dsrc28C gene. By PCR analysis, approximately equal levels of Tec transcripts containing the intact SH3 domain and containing the 22 amino acid deletion were found in hematopoietic cells. Lastly, by interspecies backcross analysis, we show that the Tec gene is tightly linked to the c-Kit gene on mouse chromosome 5.
Tec激酶最初被鉴定为一种新型细胞质蛋白酪氨酸激酶,它在肝脏中优先表达,并且与果蝇Dsrc28C src相关酪氨酸激酶高度同源。在筛选白细胞介素3(IL-3)依赖性髓系白血病细胞中的蛋白酪氨酸激酶时,我们观察到所有检测的细胞系均表达高水平的Tec转录本。然而,Tec cDNA的特征表明它们与已发表的序列有显著差异。最引人注目的是,5'区域插入41bp会影响起始密码子,并导致用94个氨基酸取代已发表的13个氨基酸的氨基末端序列。使用聚合酶链反应(PCR)分析,在造血细胞中仅检测到含有插入片段的形式。此外,我们发现一个66bp的框内插入,该插入在SH3结构域中引入了另外22个氨基酸。这种插入恢复了src基因家族和Dsrc28C基因中发现的保守SH3序列。通过PCR分析,在造血细胞中发现含有完整SH3结构域和含有22个氨基酸缺失的Tec转录本水平大致相等。最后,通过种间回交分析,我们表明Tec基因与小鼠5号染色体上的c-Kit基因紧密连锁。