Fraser M J, Tjeerde R, Matsumoto K
Can J Biochem. 1976 Nov;54(11):971-80. doi: 10.1139/o76-140.
A second form of single-strand specific endonuclease, which is stable to heating up to 74 degrees C and does not bind strongly to phosphocellulose, has been partially purified from extracts of mycelia of wild-type Neurospora crassa. The endonuclease is associated with an equally heat-stable exonuclease which degrades linear but not circular double-stranded DNA and does not attack double-stranded RNA. The exonuclease probably also degrades single-stranded DNA. Both endonuclease and exonuclease activities are inhibited by 0.1-0.5 mM ATP. The exonuclease is preferentially inhibited by a variety of agents and preferentially inactivated by trypsin. A DNA-unwinding activity has also been detected in the nuclease preparation. Protease(s) present in the nuclease preparation destroy the DNA-unwinding and exonuclease activities on incubation at 37 degrees C, but do not affect the endonuclease activity. However, the heat-stability and chromatographic properties of the endonuclease are affected by this treatment. The altered properties of the endonuclease are very similar to those of the single-strand specific endonuclease which has been previously described. The combined nuclease activities of the unaltered preparational make up a putative recombination nuclease of N. crassa.
第二种单链特异性核酸内切酶已从野生型粗糙脉孢菌的菌丝体提取物中部分纯化出来。这种核酸内切酶加热到74摄氏度仍稳定,且与磷酸纤维素结合不紧密。该核酸内切酶与一种同样耐热的核酸外切酶相关联,这种核酸外切酶可降解线性双链DNA,但不降解环状双链DNA,也不攻击双链RNA。这种核酸外切酶可能也会降解单链DNA。核酸内切酶和核酸外切酶的活性都受到0.1 - 0.5 mM ATP的抑制。这种核酸外切酶优先受到多种试剂的抑制,并优先被胰蛋白酶灭活。在核酸酶制剂中还检测到了DNA解旋活性。核酸酶制剂中存在的蛋白酶在37摄氏度孵育时会破坏DNA解旋和核酸外切酶的活性,但不影响核酸内切酶的活性。然而,这种处理会影响核酸内切酶的热稳定性和色谱性质。核酸内切酶性质的改变与先前描述的单链特异性核酸内切酶非常相似。未改变的制剂的联合核酸酶活性构成了粗糙脉孢菌一种假定的重组核酸酶。