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Translation of cellular and early vaccinia RNA in nuclease-treated lysates, derived from uninfected and vaccinia-virus-infected cells al the early stage, has been investigated. When using limiting amounts of RNA no discrimination of translation was observed in the infected cells lysates; This conclusion was confirmed by sensitive RNA competition experiments for translation in vitro and also when using two different fractionated systems for protein synthesis in vitro. This absence of detectable discrimination in vitro was established both by comparing incorporation of [35S]methionine into proteins and by analysis of the products thus synthesized by sodium dodecylsulfate gel electrophoresis. However, a modification of the translational machinery from vaccinia-virus-infected cells did occur since only the ribosomal salt wash derived from infected cells was able to reverse the inhibition of protein synthesis in vitro resulting from excess RNA (control or early). This property of vaccinia-virus-infected cell lysates may result from the synthesis of an early protein involved in translation or from a better recovery of translational factors from the infected cells., as suggested in the accompanying paper. [ABSTRACT FROM AUTHOR]

In a previous paper in this journal [Djondjurov, L., Ivanova, E. and Tsanev, R. (1979) Eur. J. Biochem. 97, 133–139], we showed that a nuclear fraction released from chromatin under a mild nuclease digestion contained an increased amount of hnRNA and the bulk of nonhistone proteins with a high metabolic rate. The present investigation has revealed that the nonhistone proteins of this fraction could be divided into three distinct metabolic groups. The first group consists of proteins with a fast turnover rate (mean half-life 30 min) which migrate into chromatin immediately after their synthesis. These proteins are predominantly acid-soluble and have relatively high molecular weights. The second group includes proteins which migrate to the nucleus more slowly and metabolize with a moderate turnover rate (mean half-life 5 h). The third group contains proteins with a more conservative metabolic behaviour. In experiments with actinomycin D it was found that the bulk of the nonhistone proteins of this fraction are not real components of the chromatin but belong to the protein moiety of heterogeneous nuclear ribonucleoprotein particles associated with chromatin. [ABSTRACT FROM AUTHOR]

When Escherichia coli 30-S ribosomal subunits are hydrolysed under mild conditions, two ribonucleoprotein fragments of unequal size are produced. Knowledge of the RNA sequences contained in these hydrolysis products was required for the experiments described in the preceding paper, and the RNA sub-fragments have therefore been examined by oligonucleotide analysis. Two well-defined small fragments of free RNA, produced concomitantly with the ribonucleoprotein fragments, were also analysed. The larger ribonucleoprotein fragment, containing predominantly proteins S4, S5, S8, S15, S16 (17) and S20, contains a complex mixture of RNA sub-fragments varying from about 100 to 800 nucleotides in length. All these fragments arose from the 5′-terminal 900 nucleotides of 16-S RNA, corresponding to the well-known 12-S fragment. No long-range interactions could be detected within this RNA region in these experiments. The RNA from the smaller ribonucleoprotein fragment (containing proteins S7, S9, S10, S14 and S19) has been described in detail previously, and consists of about 450 nucleotides near the 3′ end of the 16-S RNA, but lacking the 3′-terminal 150 nucleotides. The two small free RNA fragments (above) partly account for these missing 150 nucleotides; both fragments arose from section A of the 16-S RNA, but section J (the 3′-terminal 50 nucleotides) was not found. This result suggests that the 3′ region of 16-S RNA is not involved in stable interactions with protein. [ABSTRACT FROM AUTHOR]

1. The distribution of ribosomal particles between the soluble and membrane fractions of exponential-phase cells of Bacillus licheniformis was studied. The cells were very gently lysed by a combined treatment with lysozyme and Brij 58 in 10 mM. Tris-buffer, pH 7.6, with 60 mM K+, 10 mM Mg2+, 0.5 mM spermine and 0.75 mM spermidine. The proportion of the ribosomal material present in the membrane fraction was estimated after solubilization of the membranes by the combined action of Brij 58, deoxyribonuclease and lipase. 2. Under these conditions of lysis 96% of the total ribosomal material, consisting of subunits (12%) monosomes (20%) and polyribosomes (68%) and most of the cellular DNA, is found to be membrane-bound. 3. In the soluble fraction, containing only 4% of the total cellular amount of the ribosomal material, only subunits and 70S particles and no polyribosomes are found. 4. DNA does not seem to be involved in the binding of the ribosomal material to the membranes. 5. The results will be discussed considering the question of whether the distribution described in this paper represents the true distribution in vivo of ribosomes. [ABSTRACT FROM AUTHOR]