Fraser M J
Nucleic Acids Res. 1979 Jan;6(1):231-46. doi: 10.1093/nar/6.1.231.
Three major alkaline deoxyribonuclease (DNase) activities have been identified in sorbose-containing liquid culture medium in which wild-type Neurosporacrassa were grown: DNase A, a Ca++dependent endonuclease of molecular weight 65,000 daltons which has no specificity for single- or double-stranded DNA (ss-DNA or ds-DNA) and no activity with RNA; DNase B, a Mg++-dependent single-strand specific exonuclease of molecular weight 78,000 daltons active with both ss-DNA and RNA; DNase C, a divalent metal ion-dependent endo-exonuclease of molecular weight 65,000 having single-strand specific endonuclease activity with ss-DNA and RNA and exonuclease activity with ds-DNA. Three mutants which were shown previously to have wide spectra of sensitivities to mutagens, and which exhibited reduced release of DNase activity on sorbose-containing agar test plates (the Nuh phenotype), were deficient relative to the wild-type in the release of these major alkaline DNases into the liquid culture medium. The uvs-3 mutant released only small amounts of DNase A and DNase C; nuh-4 did not release detectable DNase C and released only a very low level of DNase B; uvs-6 released only a low level of DNase A. A nuh mutant (nuh-3) which is not mutagen sensitive relative to the wild-type released low levels of DNase B. On the other hand, an ultraviolet light-sensitive mutant (nuc-2) which does not have the Nuh phenotype was normal in the release of these DNases.
在含有山梨糖的液体培养基中培养野生型粗糙脉孢菌时,已鉴定出三种主要的碱性脱氧核糖核酸酶(DNase)活性:DNase A,一种分子量为65,000道尔顿的Ca++依赖性内切核酸酶,对单链或双链DNA(ss-DNA或ds-DNA)均无特异性,对RNA无活性;DNase B,一种分子量为78,000道尔顿的Mg++依赖性单链特异性外切核酸酶,对ss-DNA和RNA均有活性;DNase C,一种分子量为65,000的二价金属离子依赖性内切-外切核酸酶,对ss-DNA和RNA具有单链特异性内切核酸酶活性,对ds-DNA具有外切核酸酶活性。先前已证明有三种突变体对诱变剂具有广泛的敏感性谱,并且在含山梨糖的琼脂试验平板上表现出DNase活性释放减少(Nuh表型),相对于野生型,这些主要碱性DNase释放到液体培养基中的量不足。uvs-3突变体仅释放少量的DNase A和DNase C;nuh-4不释放可检测到的DNase C,仅释放极低水平的DNase B;uvs-6仅释放低水平的DNase A。相对于野生型对诱变剂不敏感的nuh突变体(nuh-3)释放低水平的DNase B。另一方面,不具有Nuh表型的紫外线敏感突变体(nuc-2)在这些DNase的释放方面是正常的。