Hoffman P D, Batschauer A, Hays J B
Department of Agricultural Chemistry, Oregon State University, Corvallis 97331-7301, USA.
Mol Gen Genet. 1996 Nov 27;253(1-2):259-65. doi: 10.1007/s004380050321.
A cDNA from Arabidopsis thaliana similar to microbial photolyase genes, and designated AT-PHH1, was isolated using a photolyase-like cDNA from Sinapsis alba (SA-PHR1) as a probe. Multiple isolations yielded only PHH1 cDNAs, and a few blue-light-receptor CRY1 (HY4) cDNAs (also similar to microbial photolyase genes), suggesting the absence of any other highly similar Arabidopsis genes. The AT-PHH1 and SA-PHR1 cDNA sequences predict 89% identity at the protein level, except for an AT-PHH1 C-terminal extension (111 amino acids), also not seen in microbial photolyases. AT-PHH1 and CRY1 show less similarity (54% p4erein identity), including respective C-terminal extensions that are themselves mostly dissimilar. Analysis of fifteen AT-PHH1 genomic isolates reveals a single gene, with three introns in the coding sequence and one in the 5'-untranslated leader. Full-length AT-PHH1, and both AT-PHH1 and AT-PHH1 delta C-513 (truncated to be approximately the size of microbial photolyase genes) cDNAs, were overexpressed, respectively, in yeast and Escherichia coli mutants hypersensitive to ultraviolet light. The absence of significant effects on resistance suggests either that any putative AT-PHH1 DNA repair activity requires cofactors/chromophores not present in yeast or E. coli, or that AT-PHH1 encodes a blue-light/ultraviolet-A receptor rather than a DNA repair protein.
以来自白芥(SA-PHR1)的类光解酶cDNA为探针,从拟南芥中分离出一个与微生物光解酶基因相似的cDNA,命名为AT-PHH1。多次分离仅得到PHH1 cDNA,以及少量蓝光受体CRY1(HY4)cDNA(也与微生物光解酶基因相似),这表明拟南芥中不存在任何其他高度相似的基因。AT-PHH1和SA-PHR1的cDNA序列在蛋白质水平上预测有89%的同一性,但AT-PHH1的C端延伸部分(111个氨基酸)除外,微生物光解酶中也未发现该延伸部分。AT-PHH1和CRY1的相似性较低(蛋白质同一性为54%),包括各自的C端延伸部分,它们本身大多不相似。对15个AT-PHH1基因组分离株的分析揭示了一个单一基因,其编码序列中有3个内含子,5'-非翻译前导序列中有1个内含子。全长AT-PHH1以及AT-PHH1和AT-PHH1 delta C-513(截短至与微生物光解酶基因大小相近)的cDNA分别在对紫外线敏感的酵母和大肠杆菌突变体中过表达。对抗性没有显著影响,这表明要么任何假定的AT-PHH1 DNA修复活性需要酵母或大肠杆菌中不存在的辅因子/发色团,要么AT-PHH1编码一种蓝光/紫外线-A受体而不是一种DNA修复蛋白。