Exo1 phosphorylation status controls the hydroxyurea sensitivity of cells lacking the Pol32 subunit of DNA polymerases delta and zeta

Exo1 belongs to the Rad2 family of structure-specific nucleases and possesses 5′–3′ exonuclease activity on double-stranded DNA substrates. Exo1 interacts physically with the DNA mismatch repair (MMR) proteins Msh2 and Mlh1 and is involved in the excision of the mispaired nucleotide. Independent of its role in MMR, Exo1 contributes to long-range resection of DNA double-strand break (DSB) ends to facilitate their repair by homologous recombination (HR), and was recently identified as a component of error-free DNA damage tolerance pathways. Here, we show that Exo1 activity increases the hydroxyurea sensitivity of cells lacking Pol32, a subunit of DNA polymerases δ and ζ. Both, phospho-mimicking and dephospho-mimicking exo1 mutants act as hypermorphs, as evidenced by an increase in HU sensitivity of pol32Δ cells, suggesting that they are trapped in an active form and that phosphorylation of Exo1 at residues S372, S567, S587, S692 is necessary, but insufficient, for the accurate regulation of Exo1 activity at stalled replication forks. In contrast, neither phosphorylation status is important for Exo1's role in MMR or in the suppression of genome instability in cells lacking Sgs1 helicase. This ability of an EXO1 deletion to suppress the HU hypersensitivity of pol32Δ cells is in contrast to the negative genetic interaction between deletions of EXO1 and POL32 in MMS-treated cells as well as the role of EXO1 in DNA-damage treated rad53 and mec1 mutants.     

Temporal dissection of an enhancer cluster reveals distinct temporal and functional contributions of individual elements

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甜苷V提取型罗汉果优良品种选育

为选育甜苷V提取型罗汉果优良品种,对19个罗汉果品种的甜苷Ⅴ含量、单株产量、果肉含量、水浸出物、总糖和总苷含量6个性状进行考察研究,综合打分评价。结果表明,F018、F014、F020综合指数较高,可作为甜苷V提取型罗汉果优良品种,为罗汉果新一代品种选育研究奠定基础。     

Mechanism of U-Insertion RNA Editing in Trypanosome Mitochondria: Characterization of RET2 Functional Domains by Mutational Analysis

3′-Terminal uridylyl transferases (TUTases) selectively bind uridine 5′-triphosphate (UTP) and catalyze the addition of uridine 5′-monophosphate to the 3′-hydroxyl of RNA substrates in a template-independent manner. RNA editing TUTase 1 and RNA editing TUTase 2 (RET2) play central roles in uridine insertion/deletion RNA editing, which is an essential part of mitochondrial RNA processing in trypanosomes. Although the conserved N-terminal (catalytic) domain and C-terminal (nucleotide base recognition) domain are readily distinguished in all known TUTases, nucleotide specificity, RNA substrate preference, processivity, quaternary structures, and auxiliary domains vary significantly among enzymes of divergent biological functions. RET2 acts as a subunit of the RNA editing core complex to carry out guide-RNA-dependent U-insertion into mitochondrial mRNA. By correlating mutational effects on RET2 activity as recombinant protein and as RNA editing core complex subunit with RNAi-based knock-in phenotypes, we have assessed the UTP and RNA binding sites in RET2. Here we demonstrate functional conservation of key UTP-binding and metal-ion-coordinating residues and identify amino acids involved in RNA substrate recognition. Invariant arginine residues 144 and 435 positioned in the vicinity of the UTP binding site are critical for RET2 activity on single-stranded and double-stranded RNAs, as well as function in vivo. Recognition of a double-stranded RNA, which resembles a guide RNA/mRNA duplex, is further facilitated by multipoint contacts across the RET2-specific middle domain.     

乙酰氨基葡萄糖转移酶Ⅴ在肿瘤转移中的作用及其分子机理

目前研究表明N-乙酰氨基葡萄糖转移酶Ⅴ在肿瘤转移中有重要作用.在恶性肿瘤中, N-乙酰氨基葡萄糖转移酶Ⅴ活性增高,其催化产物β1,6分支也增加,β1,6分支与肿 瘤的侵袭转移密切相关.本文综述了N-乙酰氨基葡萄糖转移酶Ⅴ催化形成N-糖链 β1,6分支的特点以及在N-糖链生物合成中的重要作用;还介绍了N-乙酰氨基葡萄糖转移酶Ⅴ基因组成和参与其基因调控的转录因子Ets-1,及基因表达组织特异性;着重综述了近年来N-乙酰氨基葡萄糖转移酶Ⅴ与肿瘤侵袭转移相关的分子机理的最新研究进展,包括了粘附分子钙粘蛋白(cadherin)和整合素α5β1的作用,修饰表皮生长因子受体调节信号 转导,及通过对上皮衍生的细胞表面丝氨酸蛋白酶matriptase的β1,6分支修饰促进仲瘤的 侵袭等方面.提示有效抑制N-乙酰氨基葡萄糖转移酶Ⅴ参与作用的位点,为设计抗肿瘤新药提供潜在的治疗靶点.