Flint H J
Mol Gen Genet. 1985;200(2):283-90. doi: 10.1007/BF00425437.
The effects of amino acid limitation on gene expression have been investigated in Neurospora crassa strains carrying normal (cpc-1+) or mutant (cpc-1) alleles at a locus implicated in cross-pathway amino acid control. Electrophoresis and fluorography were used to reveal the patterns of label incorporation into polypeptides in vivo, or after in vitro translation of extracted mRNAs. In a cpc-1+ strain at least 20% of detectable in vitro translation products showed relative increases in incorporation when RNA was obtained from mycelium grown under conditions of arginine limitation, by comparison with conditions of arginine sufficiency. A cpc-1 mutation, which impairs derepression of a variety of amino acid synthetic enzymes following amino acid limitation, had little detectable effect on in vivo polypeptide synthesis during amino acid sufficient growth or following pyrimidine limitation. However the mutation substantially altered the response to arginine or histidine limitation. The majority of in vitro translation products that showed increased expression in arginine limited cpc-1+ failed to increase in cpc-1 strains, but arginine limitation of cpc-1 also resulted in increases that did not occur in cpc-1+ strains. This may reflect both direct and indirect consequences of the impairment of cross-pathway control.
在粗糙脉孢菌菌株中,研究了氨基酸限制对基因表达的影响,这些菌株在一个与跨途径氨基酸控制有关的位点携带正常(cpc-1+)或突变(cpc-1)等位基因。采用电泳和荧光自显影技术来揭示体内或提取的mRNA进行体外翻译后,标记物掺入多肽的模式。在一个cpc-1+菌株中,当从在精氨酸限制条件下生长的菌丝体中获得RNA时,与精氨酸充足条件相比,至少20%可检测到的体外翻译产物显示掺入相对增加。cpc-1突变会损害氨基酸限制后多种氨基酸合成酶的去阻遏作用,在氨基酸充足生长期间或嘧啶限制后,对体内多肽合成几乎没有可检测到的影响。然而,该突变显著改变了对精氨酸或组氨酸限制的反应。在精氨酸限制的cpc-1+中显示表达增加的大多数体外翻译产物在cpc-1菌株中没有增加,但cpc-1的精氨酸限制也导致了cpc-1+菌株中未出现的增加。这可能反映了跨途径控制受损的直接和间接后果。