Messenger RNA for glutamine synthetase

Molecular and Cellular Biochemistry - Of the various eucaryotic tissues, where glutamine synthetase (GS) mRNA and its regulation have been investigated, the induction of GS by glucocorticoids in...     

Identification of Mouse Haemoglobin Messenger RNA

RETICULOCYTE polyribosomes contain 9S RNA with many of the properties expected for the haemoglobin messenger RNA (mRNA)^1–12. Proof that this RNA is the haemoglobin (Hb) mRNA, however, can be obtained only by showing that it directs the synthesis of globin chains. Laycock and Hunt¹³ added an RNA isolated from rabbit reticulocytes to an E. coli cell-free preparation and observed the synthesis of material, with the properties of globin in the presence of N-acetylvalyl tRNA. We added the mouse reticulocyte 9S RNA to a rabbit reticulocyte cell-free system and have shown that material is synthesized which co-chromatographs with mouse globin β-chains¹⁴. We now present evidence that the material synthesized under the direction of the mouse 9S RNA is indeed mouse haemoglobin β-chains.     

Translation of Globin Messenger RNA in a Heterologous Cell-free System

MESSENGER-SPECIFIC initiation factors, capable of discriminating between classes of messenger RNAs (mRNAs) or different cistrons in viral RNA, have been implicated in the regulation of protein synthesis in bacteria^1–5. Comparable but less detailed observations have also been made in eukaryotic systems^6–10. For example, RNA extracted from a mammalian virus (encephalomyocarditis virus, EMC) cannot be translated in a reticulocyte cell-free system unless the system is fortified with an extract from responsive cells—in this case, Krebs II ascites cells⁶. Such results imply the existence of tissue-specific factors and lead to questions whether this incompatibility is reciprocated by an inability of the Krebs II ascites cell system to respond to the mRNA for globin.     

Messenger RNA degradation in Saccharomyces cerevisiae

The analysis of 17 functional mRNAs and two recombinant mRNAs in the yeast Saccharomyces cerevisiae suggests that the length of an mRNA influences its half-life in this organism. The mRNAs are clearly divisible into two populations when their lengths and half-lives are compared. Differences in ribosome loading amongst the mRNAs cannot account for this division into relatively stable and unstable populations. Also, specific mRNAs seem to be destabilized to differing extents when their translation is disrupted by N-terminus-proximal stop codons. The analysis of a mutant mRNA, generated by the fusion of the yeast PYK1 and URA3 genes, suggests that a destabilizing element exists within the URA3 sequence. The presence of such elements within relatively unstable mRNAs might account for the division between the yeast mRNA populations. On the basis of these, and other previously published observations, a model is proposed for a general pathway of mRNA degradation in yeast. This model may be relevant to other eukaryotic systems. Also, only a minor extension to the model is required to explain how the stability of some eukaryotic mRNAs might be regulated.     

Messenger RNA movement on the ribosome

Recent X-ray and cryo-EM studies of 70S ribosome complexes containing different types of messenger RNAs (mRNA) and transfer RNA (tRNA) have been reviewed. Changes of the mRNA path on the ribosome at initiation and elongation states have been described. Authors suggested, that the specific region of ribosomal 30S subunit ("platform") is a ribosome binding site of regulatory domains of mRNA which locates on the non-translated 5'-end of the mRNA.     

Messenger RNA movement on the ribosome

The review summarizes the recent structural data obtained for 70S ribosome complexes with various mRNAs and tRNAs by X-ray analysis and cryoelectron microscopy. The mRNA region interacting with the ribosome at translation initiation and elongation is described. A specific part (platform) of the 30S ribosome subunit was assumed to bind the regulatory elements located in the 5′-untranslated region of mRNA.     

Messenger RNA stabilization in chicken lens development: a reexamination

Previous studies, using actinomycin D, suggested that the rate of mRNA degradation in chicken embryo lens epithelial cells decreased sharply between Days 11 and 13 of development (Yoshida and Katoh, Exp. Eye Res. 11, 1971; Exp. Cell Res. 71, 1972). We repeated these studies and directly measured the effects of actinomycin on lens mRNAs. Although actinomycin decreased [3H]uridine incorporation greater than 90%, only modest decreases in methionine incorporation were detected. Treatment for 6 hr had no detectable effect on delta-crystallin mRNA levels or on the relative amounts of proteins synthesized in an in vitro translation system. Unlike the previous studies, we did not find significant changes in the stability of lens epithelial mRNAs during development.     

Messenger RNA turnover in mouse L cells

The turnover of polyadenylic acid-containing messenger RNA and histone messenger RNA, which lacks poly(A), was studied in exponentially growing mouse L cells by measuring the kinetics of approach to steady-state uridine labeling. Constant specific activity of precursor pools was verified by showing that the data for stable RNA components, like ribosomal RNA and transfer RNA, follow theoretically predictable curves. In agreement with a previous report by Greenberg (1972), the data for poly(A)-containing mRNA (poly(A)(+)mRNA) follow theoretical curves for a class of molecules turning over with first-order (stochastic) kinetics. Cells growing with doubling times of 13·5 hours at 37 °C and 41 hours at 30 °C exhibited mean lifetimes for their poly(A)(+)mRNA of 15 hours and 42 hours, respectively, suggesting a parallelism between growth and turnover rates. The kinetic data for histone mRNA are not indicative of a stochastic process. Rather, they suggest an age-dependent decay or a zero-order (ordered) turnover with a mean lifetime of about six hours. One model, which gave a good fit to the data, considers that the histone messages persist for a fixed duration of the cell cycle, e.g. the DNA synthetic phase, and are then destroyed in a “sensitive period” after this phase. These results are discussed with regard to the possible implications of the poly(A) sequences in messenger RNA aging.     

Messenger RNA regulation in human diploid fibroblasts

In resting, non-growing human diploid fibroblasts the amount of rRNA is reduced 1.8-fold, cytoplasmic polysomes are disaggregated, and the level of poly-A RNA (mRNA) is reduced 1.8-fold in relation to growing cells. The distribution of poly-A RNA is altered in resting, non-growing cells so that an average of 64% of the total cytoplasmic poly-A RNA sediments along with particles lighter than 80S (prepolysomal) in sucrose density gradients. By comparison, in growing cells only 30% of the cytoplasmic poly-A RNA sediments in the prepolysomal region. In SDS sucrose gradients, the sedimentation profile of the prepolysomal poly-A RNA from resting cells resembles that of polysomal poly-A RNA from those cells. In contrast, the average size of prepolysomal poly-A RNA from growing cells is much smaller than that of the polysomal poly-A RNA from those cells. These data are compatible with the possibility that resting cell prepolysomal poly-A is untranslated mRNA. Also consistent with this interpretation are experiments which demonstrate that one-quarter to one-third of the prepolysomal poly-A RNA of resting cells is recruited into polysomes in the presence of cycloheximide.