Hoenicka Janet, Fernández Lobato María, Marín Dolores, Jiménez Antonio
Centro de Biología Molecular, Universidad Autónoma de Madrid, Canto Blanco, 28049 Madrid, Spain.
Yeast. 2002 Jun 30;19(9):735-43. doi: 10.1002/yea.871.
In Saccharomyces cerevisiae, the SCR1 gene from Schwanniomyces occidentalis is known to induce ribosomal resistance to cycloheximide (cyh). A 2.8 kb DNA fragment encoding this gene was sequenced. Its EMBL Accession No. is AJ419770. It disclosed a putative tRNA(Asn) (GUU) sequence located downstream of an open reading frame (ORF) of 1641 nucleotides. This ORF was shown to correspond to SCR1. It would encode a highly hydrophobic polypeptide (SCR1) with 12 transmembrane domains. SCR1 is highly similar to a variety of yeast proteins of the multidrug-resistance (MDR) family. However, SCR1 only conferred resistance to cyh but not to benomyl or methotrexate. The cyh-resistance phenotype induced by SCR1 was confirmed in several S. cerevisiae strains that expressed this gene to reside at the ribosomal level. In contrast, a beta-galacosidase-tagged SCR1 was found to be integrated in the endoplasmic reticulum (ER). It is proposed that the ribosomes of yeast cells expressing SCR1 undergo a conformational change during their interaction with the ER, which lowers their affinity for cyh-binding. If so, these findings would disclose a novel ribosomal resistance mechanism.
在酿酒酵母中,已知西方施万酵母的SCR1基因可诱导核糖体对环己酰亚胺(cyh)产生抗性。对编码该基因的一个2.8 kb DNA片段进行了测序。其EMBL登录号为AJ419770。该片段揭示了一个推定的tRNA(Asn)(GUU)序列,位于一个1641个核苷酸的开放阅读框(ORF)下游。该ORF被证明对应于SCR1。它将编码一个具有12个跨膜结构域的高度疏水多肽(SCR1)。SCR1与多种多药耐药(MDR)家族的酵母蛋白高度相似。然而,SCR1仅赋予对cyh的抗性,而不赋予对苯菌灵或甲氨蝶呤的抗性。在几个表达该基因的酿酒酵母菌株中,证实了SCR1诱导的cyh抗性表型存在于核糖体水平。相反,发现一个β-半乳糖苷酶标记的SCR1整合在内质网(ER)中。有人提出,表达SCR1的酵母细胞核糖体在与ER相互作用期间会发生构象变化,这降低了它们对cyh结合的亲和力。如果是这样,这些发现将揭示一种新的核糖体抗性机制。