Department of Molecular Biology, University of Geneva, Switzerland.
Carcinogenesis. 2012 Jun;33(6):1203-10. doi: 10.1093/carcin/bgs149. Epub 2012 Apr 16.
The vertebrate RECQL4 (RECQ4) gene is thought to be the ortholog of budding yeast SLD2. However, RecQL4 contains within its C-terminus a RecQ-like helicase domain, which is absent in Sld2. We established human pre-B lymphocyte Nalm-6 cells, in which the endogenous RECQL4 gene was homozygously targeted such that the entire C-terminus would not be expressed. The RECQL4(ΔC/ΔC) cells behaved like the parental cells during unperturbed DNA replication or after treatment with agents that induce stalling of DNA replication forks, such as hydroxyurea (HU). However, after exposure to ionizing radiation (IR), the RECQL4(ΔC/ΔC) cells exhibited hypersensitivity, inability to complete S phase and prematurely terminated or paused DNA replication forks. Deletion of BLM, a gene that also encodes a RecQ helicase, had the opposite phenotype; an almost wild-type response to IR, but hypersensitivity to HU. Targeting both RECQL4 and BLM resulted in viable cells, which exhibited mostly additive phenotypes compared with those exhibited by the RECQL4(ΔC/ΔC) and the BLM(-/-) cells. We propose that RecQL4 facilitates DNA replication in cells that have been exposed to IR.
脊椎动物 RECQL4(RECQ4)基因被认为是芽殖酵母 SLD2 的直系同源物。然而,RecQL4 在其 C 末端包含一个 RecQ 样解旋酶结构域,而 Sld2 中则没有。我们建立了人前 B 淋巴细胞 Nalm-6 细胞,其中内源性 RECQL4 基因被纯合靶向,使得整个 C 末端不会表达。在未受干扰的 DNA 复制期间或在用诱导 DNA 复制叉停滞的试剂(如羟基脲(HU))处理后,RECQL4(ΔC/ΔC)细胞的行为与亲本细胞相似。然而,暴露于电离辐射(IR)后,RECQL4(ΔC/ΔC)细胞表现出敏感性增加、无法完成 S 期以及过早终止或暂停 DNA 复制叉。BLM 基因的缺失,该基因也编码一种 RecQ 解旋酶,表现出相反的表型;对 IR 的反应几乎与野生型相同,但对 HU 敏感。靶向 RECQL4 和 BLM 都会导致存活细胞,与 RECQL4(ΔC/ΔC)和 BLM(-/-)细胞相比,这些细胞表现出主要的加性表型。我们提出 RecQL4 有助于细胞在暴露于 IR 后进行 DNA 复制。