Zanotti Kimberly J, Maul Robert W, Castiblanco Diana P, Yang William, Choi Yong Jun, Fox Jennifer T, Myung Kyungjae, Saribasak Huseyin, Gearhart Patricia J
Laboratory of Molecular Biology and Immunology, National Institute on Aging, National Institutes of Health, Baltimore, MD 21224; and.
Genetics and Molecular Biology Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892.
J Immunol. 2015 Jan 1;194(1):35-42. doi: 10.4049/jimmunol.1401158. Epub 2014 Nov 17.
Mammalian ATPase family AAA domain-containing protein 5 (ATAD5) and its yeast homolog enhanced level of genomic instability 1 are responsible for unloading proliferating cell nuclear antigen from newly synthesized DNA. Prior work in HeLa and yeast cells showed that a decrease in ATAD5 protein levels resulted in accumulation of chromatin-bound proliferating cell nuclear antigen, slowed cell division, and increased genomic instability. In this study, B cells from heterozygous (Atad5(+/m)) mice were used to examine the effects of decreased cell proliferation on Ab diversity. ATAD5 haploinsufficiency did not change the frequency or spectrum of somatic hypermutation in Ab genes, indicating that DNA repair and error-prone DNA polymerase η usage were unaffected. However, immunized Atad5(+/m) mice had decreased serum IgG1 Abs, demonstrating a functional effect on class switch recombination. The mechanism of this altered immune response was then examined following ex vivo stimulation of splenic B cells, where Atad5(+/m) cells accumulated in the S phase of the cell cycle and had reduced proliferation compared with wild-type cells. These haploinsufficient cells underwent a significant decline in activation-induced deaminase expression, resulting in decreased switch region DNA double-strand breaks and interchromosomal translocations in the Igh locus. Class switch recombination to several isotypes was also reduced in Atad5(+/m) cells, although the types of end-joining pathways were not affected. These results describe a defect in DNA replication that affects Igh recombination via reduced cell division.
哺乳动物含ATP酶家族AAA结构域蛋白5(ATAD5)及其酵母同源物基因组不稳定增强因子1负责从新合成的DNA上卸载增殖细胞核抗原。先前在HeLa细胞和酵母细胞中的研究表明,ATAD5蛋白水平降低会导致染色质结合的增殖细胞核抗原积累、细胞分裂减慢以及基因组不稳定增加。在本研究中,利用杂合(Atad5(+/m))小鼠的B细胞来检测细胞增殖减少对抗体多样性的影响。ATAD5单倍体不足并未改变抗体基因中体细胞高频突变的频率或谱型,这表明DNA修复和易出错的DNA聚合酶η的使用未受影响。然而,免疫后的Atad5(+/m)小鼠血清IgG1抗体减少,这表明对类别转换重组有功能影响。随后,在体外刺激脾B细胞后,研究了这种改变的免疫反应机制,其中Atad5(+/m)细胞在细胞周期的S期积累,与野生型细胞相比增殖减少。这些单倍体不足的细胞中激活诱导的脱氨酶表达显著下降,导致转换区DNA双链断裂减少以及Igh基因座中的染色体间易位减少。Atad5(+/m)细胞中向几种同种型的类别转换重组也减少,尽管末端连接途径的类型未受影响。这些结果描述了一种DNA复制缺陷,其通过减少细胞分裂影响Igh重组。