Johnson R E, Washington M T, Prakash S, Prakash L
Sealy Center for Molecular Science, University of Texas Medical Branch, Galveston, Texas 77555-1061, USA.
J Biol Chem. 2000 Mar 17;275(11):7447-50. doi: 10.1074/jbc.275.11.7447.
Xeroderma pigmentosum (XP) patients are highly sensitive to sunlight, and they suffer from a high incidence of skin cancers. The variant form of XP results from mutations in the hRAD30A gene, which encodes the DNA polymerase in humans, hPol(eta). Of the eukaryotic DNA polymerases, only human Pol(eta) and its yeast counterpart have the ability to replicate DNA containing a cis-syn thymine-thymine (T-T) dimer. Here we measure the fidelity of hPol(eta) on all four nondamaged template bases and at each thymine residue of a cis-syn T-T dimer. Opposite all four nondamaged template bases, hPol(eta) misincorporates nucleotides with a frequency of approximately 10(-2)-10(-3), and importantly, hPol(eta) synthesizes DNA opposite the T-T dimer with the same accuracy and efficiency as opposite the nondamaged DNA. The low fidelity of hPol(eta) may derive from a flexible active site that renders the enzyme more tolerant of geometric distortions in DNA and enables it to synthesize DNA past a T-T dimer.
着色性干皮病(XP)患者对阳光高度敏感,且患皮肤癌的几率很高。XP的变异形式是由hRAD30A基因突变引起的,该基因编码人类DNA聚合酶hPol(η)。在真核生物DNA聚合酶中,只有人类Pol(η)及其酵母对应物有能力复制含有顺式-胸腺嘧啶-胸腺嘧啶(T-T)二聚体的DNA。在此,我们测定了hPol(η)在所有四种未受损模板碱基上以及顺式-胸腺嘧啶-胸腺嘧啶二聚体每个胸腺嘧啶残基处的保真度。在所有四种未受损模板碱基对面,hPol(η)错误掺入核苷酸的频率约为10^(-2)-10^(-3),重要的是,hPol(η)与未受损DNA对面相比,以相同的准确性和效率合成T-T二聚体对面的DNA。hPol(η)的低保真度可能源于一个灵活的活性位点,该位点使酶对DNA中的几何扭曲更具耐受性,并使其能够在T-T二聚体后合成DNA。