期刊
NUCLEIC ACIDS RESEARCH
卷 40, 期 14, 页码 6608-6619出版社
OXFORD UNIV PRESS
DOI: 10.1093/nar/gks318
关键词
-
资金
- Medical Research Council [G0600776]
- Cancer Research UK [C6563/A10192]
- Medical Research Council
- MRC [MR/J006750/1, G0600776] Funding Source: UKRI
- Medical Research Council [G0801130B, MR/J006750/1, G0600776] Funding Source: researchfish
Microcephaly with early-onset, intractable seizures and developmental delay (MCSZ) is a hereditary disease caused by mutations in polynucleotide kinase/phosphatase (PNKP), a DNA strand break repair protein with DNA 5'-kinase and DNA 3'-phosphatase activity. To investigate the molecular basis of this disease, we examined the impact of MCSZ mutations on PNKP activity in vitro and in cells. Three of the four mutations currently associated with MCSZ greatly reduce or ablate DNA kinase activity of recombinant PNKP at 30 degrees C (L176F, T424Gfs48X and exon15 delta fs4X), but only one of these mutations reduces DNA phosphatase activity under the same conditions (L176F). The fourth mutation (E326K) has little impact on either DNA kinase or DNA phosphatase activity at 30 degrees C, but is less stable than the wild-type enzyme at physiological temperature. Critically, all of the MCSZ mutations identified to date result in similar to 10-fold reduced cellular levels of PNKP protein, and reduced rates of chromosomal DNA strand break repair. Together, these data suggest that all four known MCSZ mutations reduce the cellular stability and level of PNKP protein, with three mutations likely ablating cellular DNA 5'-kinase activity and all of the mutations greatly reducing cellular DNA 3'-phosphatase activity.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据