Margulies M M, Michaels A
J Cell Biol. 1974 Jan;60(1):65-77. doi: 10.1083/jcb.60.1.65.
The amount of chloroplast ribosomal RNAs of Chlamydomonas reinhardtii which sediment at 15,000 g is increased when cells are treated with chloramphenicol. Preparations of chloroplast membranes from chloramphenicol-treated cells contain more chloroplast ribosomal RNAs than preparations from untreated cells. The membranes from treated cells also contain more ribosome-like particles, some of which appear in polysome-like arrangements. About 50% of chloroplast ribosomes are released from membranes in vitro as subunits by 1 mM puromycin in 500 mM KCl. A portion of chloroplast ribosomal subunits is released by 500 mM KCl alone, a portion by 1 mM puromycin alone, and a portion by 1 mM puromycin in 500 mM KCl. Ribosomes are not released from isolated membranes by treatment with ribonuclease. Membranes in chloroplasts of chloramphenicol-treated cells show many ribosomes associated with membranes, some of which are present in polysome-like arrangements. This type of organization is less frequent in chloroplasts of untreated cells. Streptogramin, an inhibitor of initiation, prevents chloramphenicol from acting to permit isolation of membrane-bound ribosomes. Membrane-bound chloroplast ribosomes are probably a normal component of actively growing cells. The ability to isolate membrane-bound ribosomes from chloramphenicol-treated cells is probably due to chloramphenicol-prevented completion of nascent chains during harvesting of cells. Since chloroplasts synthesize some of their membrane proteins, and a portion of chloroplast ribosomes is bound to chloroplast membranes through nascent protein chains, it is suggested that the membrane-bound ribosomes are synthesizing membrane protein.
用氯霉素处理莱茵衣藻细胞后,在15,000 g下沉降的叶绿体核糖体RNA的量会增加。与未处理细胞的叶绿体膜制备物相比,经氯霉素处理细胞的叶绿体膜制备物含有更多的叶绿体核糖体RNA。处理后细胞的膜中还含有更多核糖体样颗粒,其中一些呈多核糖体样排列。在体外,1 mM嘌呤霉素在500 mM KCl中可使约50%的叶绿体核糖体以亚基形式从膜上释放。一部分叶绿体核糖体亚基仅由500 mM KCl释放,一部分仅由1 mM嘌呤霉素释放,还有一部分由1 mM嘌呤霉素在500 mM KCl中释放。用核糖核酸酶处理不能使核糖体从分离的膜上释放。经氯霉素处理细胞的叶绿体中的膜显示许多核糖体与膜结合,其中一些呈多核糖体样排列。这种组织类型在未处理细胞的叶绿体中较少见。起始抑制剂链阳霉素可阻止氯霉素发挥作用以允许分离膜结合核糖体。膜结合的叶绿体核糖体可能是活跃生长细胞的正常组成部分。从经氯霉素处理的细胞中分离膜结合核糖体的能力可能是由于在收获细胞期间氯霉素阻止了新生链的完成。由于叶绿体合成其一些膜蛋白,并且一部分叶绿体核糖体通过新生蛋白链与叶绿体膜结合,因此有人提出膜结合核糖体正在合成膜蛋白。