Murine Cell Line SX9 Bearing a Mutation in the dna-pkcsGene Exhibits Aberrant V(D)J Recombination Not Only in the Coding Joint but Also in the Signal Joint*

Murine Cell Line SX9 Bearing a Mutation in the dna-pkcsGene Exhibits Aberrant V(D)J Recombination Not Only in the Coding Joint but Also in the Signal Joint*
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带有 dna-pkcs 基因突变的鼠细胞系 SX9 不仅在编码接头中而且在信号接头中都表现出异常的 V(D)J 重组*

DOI:
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发表时间:
1998
影响因子:
4.8
通讯作者:
M. Abe
M. Abe
中科院分区:
生物学2区
文献类型:
--
作者:
R. Fukumura;R. Araki;A. Fujimori;M. Mori;Toshiyuki Saito;Fumiaki Watanabe;Mika Sarashi;H. Itsukaichi;K. Eguchi;Koki Sato;K. Tatsumi;M. Abe

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我们从乳腺癌细胞系FM3A中建立了放射敏感细胞系SX9。SX9细胞存在dna依赖性蛋白激酶(DNA-PK)活性缺陷。此外,互补试验表明,SX9细胞系属于x射线交叉互补组(XRCC) 7。SX9细胞中dna依赖性蛋白激酶催化亚基(dna-pkcs) cDNA的分离和序列分析显示核苷酸“T”(9572)到“C”的转变导致氨基酸残基亮氨酸(3191)被脯氨酸取代。有趣的是,突变发生在一个等位基因上,而dna-pkcs的转录本完全来自突变的等位基因。利用染色体外载体进行V(D)J重组试验,不仅发现了编码缺陷,还发现了信号关节形成缺陷。与正常细胞系相比,信号接头的频率下降到大约十分之一,保真度急剧下降到12.2%。为了确认dna-pkcs基因对SX9中V(D)J异常重组的作用,我们将全长dna-pkcs基因引入SX9。结果,观察到野生型dna-pkcs cDNA对V(D)J重组的恢复。SX9是一种新颖的dna-pkcs缺陷细胞系。
We established the radiosensitive cell line SX9 from mammary carcinoma cell line FM3A. In SX9 cells a defect of DNA-dependent protein kinase (DNA-PK) activity was suggested. Additionally, a complementation test suggested that the SX9 cell line belongs to a x-ray cross-complementing group (XRCC) 7. Isolation and sequence analyses of DNA-dependent protein kinase catalytic subunit (dna-pkcs) cDNA in SX9 cells disclosed nucleotide “T” (9572) to “C” transition causing substitution of amino acid residue leucine (3191) to proline. Interestingly, the mutation occurs in one allele, and transcripts of the dna-pkcs expressed exclusively from mutated allele. V(D)J recombination assay using extrachromosomal vector revealed the defects of not only coding but also signal joint formation. The frequency of the signal joint decreased to approximately one-tenth and the fidelity drastically decreased to 12.2% as compared with the normal cell line. To confirm the responsibility of thedna-pkcs gene for abnormal V(D)J recombination in SX9, the full-length dna-pkcs gene was introduced into SX9. As a result, restoration of V(D)J recombination by wild typedna-pkcs cDNA was observed. SX9 is a noveldna-pkcs-deficient cell line.
DOI: --
发表时间: 1997
期刊: Radiation research
影响因子: 3.4
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xrs-6 细胞系中的 V(D)J 重组缺陷是由 Ku80 基因的点突变引起的。
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马 SCID 中 DNA-PK(CS) 的激酶阴性突变会导致编码和信号接头形成缺陷。
DOI: --
发表时间: 1997
期刊: Journal of immunology (Baltimore, Md. : 1950)
影响因子: --
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通讯作者: Meek,K
DOI: 10.1126/science.8073286
发表时间: 1994-09-02
期刊: SCIENCE
影响因子: 56.9
作者:
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通讯作者: JEGGO, PA
DOI: 10.1073/pnas.92.25.11485
发表时间: 1995-12-05
影响因子: 11.1
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WILER, R;LEBER, R;MEEK, K
通讯作者: MEEK, K