Taylor M W, Pipkorn J H, Tokito M K, Pozzatti R O
Somatic Cell Genet. 1977 Mar;3(2):195-206. doi: 10.1007/BF01551814.
Spontaneous and mutagen-induced 2,6-diaminopurine-resistant mutants of Chinese hamster ovary (CHO-K1) cells were isolated. Such mutants fell into two classes: spontaneous and ethylmethane-sulfonate-induced mutants had approximately 5% wild-type adenine phosphoribosyl transferase (APRT) activity, whereas ICR-170G-induced mutants had barely detectable APRT activity. Since it has been reported that human hypoxanthine-guanine phosphoribosyl transferase (HGPRT) (Lesch-Nyhan syndrome) and APRT mutants over-produce purines, we examined the control and rate of purine biosynthesis in the Chinese hamster mutants. End product inhibition by adenine could not be demonstrated in such mutants, indicating that the active feedback inhibitor is a nucleotide rather than the free purine base, HGPRT activity was normal in all mutants examined except in one isolate. Purine biosynthesis as measured by the accumulation of the purine biosynthetic intermediate phosphoribosyl formylglycineamide was not elevated in the mutants as might have been predicted from work with Lesch-Nyhan cells. The data also suggest that our strain of CHO-K1 is physically or functionally haploid for the APRT locus.
分离出了中国仓鼠卵巢(CHO-K1)细胞的自发及诱变剂诱导的抗2,6-二氨基嘌呤突变体。这些突变体分为两类:自发突变体和经甲基磺酸乙酯诱导的突变体具有约5%的野生型腺嘌呤磷酸核糖转移酶(APRT)活性,而经ICR-170G诱导的突变体几乎检测不到APRT活性。由于已有报道称人类次黄嘌呤-鸟嘌呤磷酸核糖转移酶(HGPRT)(莱施-奈恩综合征)和APRT突变体会过量产生嘌呤,我们检测了中国仓鼠突变体中嘌呤生物合成的调控及速率。在这些突变体中未证实腺嘌呤对终产物的抑制作用,这表明活性反馈抑制剂是一种核苷酸而非游离嘌呤碱,在所检测的所有突变体中,除一个分离株外,HGPRT活性均正常。通过嘌呤生物合成中间产物磷酸核糖甲酰甘氨酰胺的积累来衡量,突变体中的嘌呤生物合成并未如对莱施-奈恩细胞的研究预期那样升高。数据还表明,我们的CHO-K1细胞株在APRT基因座上在物理或功能上是单倍体。