scholarly journals Regulation of polyribosome formation and protein synthesis in the uterus. Effect of ovariectomy and administration of oestradiol-17β on the amino acid-incorporation activity in vitro and the cytoplasmic concentration in vivo of polyribosomal preparation

1967 ◽  
Vol 105 (3) ◽  
pp. 1101-1109 ◽  
Author(s):  
Ching-Sung Teng ◽  
Terrell H. Hamilton
Keyword(s):  
Protein Synthesis ◽  
Amino Acid ◽  
Hormonal Regulation ◽  
Ovariectomized Rats ◽  
Cytoplasmic Protein ◽  
Amino Acid Incorporation ◽  
Free System ◽  
Acid Incorporation

1. The hormonal regulation of cytoplasmic protein synthesis in the uterus is described. Polyribosomal preparation from uteri of normal or ovariectomized rats was isolated by procedure 3 and assayed for [14C]leucine-incorporation activity in the cell-free system, as described by Teng & Hamilton (1967). 2. Ovariectomy of normal animals caused, 3 weeks after surgery, a 50–60% increase in the amino acid-incorporation activity in vitro of uterine polyribosomal preparation, but a 90–95% decrease in the cytoplasmic concentration in vivo of the preparation. 3. Administration of 10μg. of oestradiol-17β to ovariectomized rats at zero time caused, 10–12hr. later, a 100% stimulation in amino acid-incorporation activity in vitro of the uterine polyribosomal preparation. From 12hr. to 36hr. after hormone administration, the activity in vitro of the preparation decreased. If a second dose of hormone was administered at 36hr., the activity in vitro of the preparation continued to decrease, and approached at 48hr. and 72hr. the lower activity observed for the preparation from normal animals. The cytoplasmic concentration of polyribosomal preparation increased by 600–700% under these experimental conditions. If a second dose of oestradiol-17β was not administered at 36hr., the initially elevated cytoplasmic concentration of the preparation decreased by 50% from 36hr. to 72hr., and the activity in vitro of the preparation was not fully depressed to the ‘normal’ value. 4. Pretreatment of ovariectomized animals with actinomycin D or cycloheximide abolished 80–90% of the stimulatory effects of hormone treatment on the amino acid-incorporation activity in vitro and cytoplasmic concentration in vivo of uterine polyribosomal preparation. 5. Two major conclusions are drawn from the results reported: that during early oestrogen action new polyribosomes having amino acid-incorporation properties different from those of the old ones appear and accumulate in the cytoplasm of the uterus; and that the regulation of cytoplasmic protein synthesis in the organ by oestrogen is of an indirect nature, with dual effects of the hormone on genetic transcription resulting in turn in a regulation of the rate and amount of genetic translation.

Effect of Cycloheximide on In Vitro and In Vivo Protein Synthesis by Certain Tissues of Rats and Pigeons with Special Reference to Muscle

1973 ◽  
Vol 51 (12) ◽  
pp. 933-941 ◽  
Author(s):  
Njanoor Narayanan ◽  
Jacob Eapen
Keyword(s):  
Amino Acids ◽  
Body Weight ◽  
Protein Synthesis ◽  
Amino Acid ◽  
Amino Acid Incorporation ◽  
Contrast Administration ◽  
Acid Incorporation ◽  
Tissue Homogenates

The effect of cycloheximide in vitro and in vivo on the incorporation of labelled amino acids into protein by muscles, liver, kidneys, and brain of rats and pigeons was studied. In vitro incorporation of amino acids into protein by muscle microsomes, myofibrils, and myofibrillar ribosomes was not affected by cycloheximide. In contrast, administration of the antibiotic into intact animals at a concentration of 1 mg/kg body weight resulted in considerable inhibition of amino acid incorporation into protein by muscles, liver, kidneys, and brain. This inhibition was observed in all the subcellular fractions of these tissues during a period of 10–40 min after the administration of the precursor. Tissue homogenates derived from in vivo cycloheximide-treated animals did not show significant alteration in in vitro amino acid incorporation with the exception of brain, which showed a small but significant enhancement.

scholarly journals Biotin-mediated protein biosynthesis

1974 ◽  
Vol 140 (3) ◽  
pp. 549-556 ◽  
Author(s):  
R. L. Boeckx ◽  
K. Dakshinamurti
Keyword(s):  
Protein Synthesis ◽  
Amino Acid ◽  
Rat Liver ◽  
Intestinal Mucosa ◽  
Protein Biosynthesis ◽  
Amino Acid Incorporation ◽  
Acid Incorporation ◽  
Stimulation Of

The effect of administration of biotin to biotin-deficient rats on protein biosynthesis was studied. Biotin treatment resulted in stimulation by more than twofold of amino acid incorporation into protein, both in vivo and in vitro in rat liver, pancreas, intestinal mucosa and skin. Analysis of the products of amino acid incorporation into liver proteins in vivo and in vitro indicated that the synthesis of some proteins was stimulated more than twofold, but others were not stimulated at all. This indicates a specificity in the stimulation of protein synthesis mediated by biotin.

THYROXINE STIMULATION OF AMINO ACID INCORPORATION INTO MITOCHONDRIAL PROTEIN: DIFFERENCES BETWEEN IN VIVO AND IN VITRO EFFECTS

1973 ◽  
Vol 72 (4) ◽  
pp. 684-696 ◽  
Author(s):  
Amirav Gordon ◽  
Martin I. Surks ◽  
Jack H. Oppenheimer
Keyword(s):  
Protein Synthesis ◽  
Amino Acid ◽  
Mitochondrial Protein ◽  
Leucine Incorporation ◽  
Mitochondrial Protein Synthesis ◽  
Amino Acid Incorporation ◽  
Acid Incorporation ◽  
Stimulation Of

ABSTRACT The in vivo and in vitro stimulation of rat hepatic mitochondrial protein synthesis by thyroxine (T4) was compared. In confirmation of Buchanan & Tapley (1966). T4 added to isolated mitochondria rapidly stimulated [14C] leucine incorporation into mitochondrial protein. The in vitro stimulation was reversed after T4 was removed by incubating the mitochondria with bovine serum albumin (BSA). The decrease in T4 stimulation of protein synthesis appeared proportional to the T4 removed by BSA. Thus, it appears probable that exchangeable T4 controls the in vitro system. In contrast, the increase in mitochondrial protein synthesis which was observed 3 to 4 days after pretreatment of hypothyroid rats with labelled and non-radioactive T4 was not reversed by BSA treatment. Moreover, mitochondrial radioactivity could not be extracted with albumin. The in vivo phenomenon does not, therefore, appear to be related to exchangeable hormone in the mitochondria. Furthermore, the estimated quantity of T4 associated with mitochondria after in vivo stimulation was at least two orders of magnitude less than that required to produce comparable stimulation of mitochondrial protein synthesis in vitro. These findings strongly suggest that in vitro and in vivo stimulation of amino acid incorporation by T4 may be mediated by different biochemical mechanisms.

Suppressors suaC 109 and suaA 101 of Aspergillus nidulans alter the ribosomal phenotype in vitro

1987 ◽  
Vol 7 (12) ◽  
pp. 941-948 ◽  
Author(s):  
A. Zamir ◽  
S. S. Martinelli
Keyword(s):  
Protein Synthesis ◽  
Amino Acid ◽  
Aspergillus Nidulans ◽  
Ribosomal Proteins ◽  
Amino Acid Incorporation ◽  
Free System ◽  
Cell Free System ◽  
Translation Factors ◽  
Acid Incorporation

A new homologous, cell-free system for protein synthesis has been devised for use with ribosomes and elongation factors from Aspergillus nidulans. Ribosome preparations from strains with either the suaAlO1 or suaCl09 mutations have a higher misreading ratio (non-cognate:cognate amino acid incorporation) in the presence of hygromycin than controls. They can be classed as fidelity mutants. These results also prove that the mutations must be in genes coding for ribosomal proteins or enzymes which modify ribosomal proteins post-translationally. Alternatively, the genes could code for translation factors.

An in vitro amino acid incorporation method for assessing the status of in vivo protein synthesis

1984 ◽  
Vol 136 (2) ◽  
pp. 285-292 ◽  
Author(s):  
Thaddeus S. Nowak ◽  
Elizabeth R. Carty ◽  
W.David Lust ◽  
Janet V. Passonneau
Keyword(s):  
Protein Synthesis ◽  
Amino Acid ◽  
Amino Acid Incorporation ◽  
Acid Incorporation ◽  
The Status ◽  
Incorporation Method ◽  
In Vivo Protein Synthesis

scholarly journals Regulation of polyribosome formation and protein synthesis in the uterus. Effect of administration of graded doses of oestradiol-17β and actinomycin D on the amino acid-incorporation activity in vitro and the cytoplasmic concentration in vivo of polyribosomes

1970 ◽  
Vol 118 (3) ◽  
pp. 341-346 ◽  
Author(s):  
Ching-Sung Teng ◽  
Terrell H. Hamilton
Keyword(s):  
Amino Acid ◽  
Molecular Mechanisms ◽  
Actinomycin D ◽  
Amino Acid Incorporation ◽  
Free System ◽  
Acid Incorporation ◽  
Minimal Dose ◽  
Early Action

1. The effects of graded doses of oestradiol-17β and actinomycin D, administered separately or together, on the amino acid-incorporation activity in vitro and the cytoplasmic concentration in vivo of uterine polyribosomes are described. Preparations of polyribosomes isolated from uteri of ovariectomized adult rats were determined for cytoplasmic concentration in vivo and assayed for [14C]leucine-incorporation activity in the cell-free system, exactly as described by Teng & Hamilton (1967b). 2. A minimal dose of 10μg of oestradiol-17β administered for 10h was found to increase, by about 100%, both the amino acid-incorporation activity in vitro and the cytoplasmic concentration in vivo of the polyribosomes. A minimal dose of 250μg of actinomycin D administered for 10h was found to inhibit, by about 50%, the incorporation activity in vitro of the polyribosomes. All doses of the inhibitor administered for 10h failed to alter the cytoplasmic concentration in vivo of the polyribosomes. 3. A dose of 10μg of oestradiol-17β restored to the control value the inhibitory effect of a dose of either 50 or 125μg of actinomycin D on the activity in vitro of the polyribosomes, at 10h after treatment with the inhibitor and the hormone. In these experiments, there was an increase of 60–100% in the cytoplasmic concentration in vivo of the polyribosomes. 4. A dose of 125μg of actinomycin D, administered to animals along with 10μg of oestradiol-17β for 6–36h, abolished the hormone-induced enhancement of the incorporation activity in vitro, but did not prevent an increase of about 200% in the cytoplasmic concentration in vivo of the polyribosomes. However, treatment with 750μg of the inhibitor abolished both stimulatory effects of the hormone. 5. The results reported indicate that the stimulatory effects of oestradiol-17β in vivo on the number and activity of the cytoplasmic polyribosomes in the uterus of the ovariectomized rat have different sensitivities to actinomycin D, but the primary molecular mechanisms responsible for the results are unknown. The major conclusion drawn is that the formation and appearance in the cytoplasm of newly formed polyribosomes are important features of the early action of oestrogen in the uterus.

Reduced and S-carboxymethylated human growth hormone: a probe for diabetogenic action

1984 ◽  
Vol 247 (5) ◽  
pp. E639-E644
Author(s):  
C. M. Cameron ◽  
J. L. Kostyo ◽  
J. A. Rillema ◽  
S. E. Gennick
Keyword(s):  
Growth Hormone ◽  
Amino Acid ◽  
Human Growth Hormone ◽  
Human Growth ◽  
Mouse Mammary Gland ◽  
Amino Acid Incorporation ◽  
Acid Incorporation ◽  
Hypophysectomized Rats

The biological activity profile of reduced and S-carboxymethylated human growth hormone (RCM-hGH) was determined to establish its suitability for study of the diabetogenic property of hGH. RCM-hGH was found to have greatly attenuated in vivo growth-promoting activity in the 9-day weight-gain test in hypophysectomized rats (approximately 1%) and to have a similar low order of in vitro activity in stimulating amino acid incorporation into the protein of the isolated rat diaphragm. RCM-hGH also only had approximately 1% of the in vitro insulin-like activity of the native hormone on isolated adipose tissue from hypophysectomized rats. In contrast, RCM-hGH retained substantial in vivo diabetogenic activity in the ob/ob mouse, appearing to have approximately 50% of the activity of the native hormone. RCM-hGH was also found to retain significant, although attenuated (25%), in vitro lactogenic activity when tested for the ability to stimulate amino acid incorporation into a casein-rich protein fraction in mouse mammary gland explants. Because RCM-hGH exhibits a high degree of diabetogenic activity, although lacking significant anabolic or insulin-like activities, it will be useful as a "monovalent" probe for the study of the molecular mechanism of the diabetogenic action of GH.

scholarly journals Action of growth hormone in vitro on the net uptake and incorporation into protein of amino acids in muscle from intact rabbits given protein-deficient diets

1976 ◽  
Vol 35 (1) ◽  
pp. 1-10 ◽  
Author(s):  
M. R. Turner ◽  
P. J. Reeds ◽  
K. A. Munday
Keyword(s):  
Amino Acids ◽  
Growth Hormone ◽  
Protein Synthesis ◽  
Amino Acid ◽  
De Novo ◽  
Amino Acid Uptake ◽  
Acid Uptake ◽  
Amino Acid Incorporation ◽  
Acid Incorporation

1. Net amino acid uptake, and incorporation into protein have been measured in vitro in the presence and absence of porcine growth hormone (GH) in muscle from intact rabbits fed for 5 d on low-protein (LP), protein-free (PF) or control diets.2. In muscle from control and LP animals GH had no effect on the net amino acid uptake but stimulated amino acid incorporation into protein, although this response was less in LP animals than in control animals.3. In muscle from PF animals, GH stimulated both amino acid incorporation into protein and the net amino acid uptake, a type of response which also occurs in hypophysectomized animals. The magnitude of the effect of GH on the incorporation of amino acids into protein was reduced in muscle from PF animals.4. The effect of GH on the net amino acid uptake in PF animals was completely blocked by cycloheximide; the uptake effect of GH in these animals was dependent therefore on de novo protein synthesis.5. It is proposed that in the adult the role of growth hormone in protein metabolism is to sustain cellular protein synthesis when there is a decrease in the level of substrate amino acids, similar to that which occurs during a short-term fast or when the dietary protein intake is inadequate.

An in vitro bioassay for a mulluscan gonadotropin

1975 ◽  
Vol 62 (2) ◽  
pp. 433-446
Author(s):  
M. J. Wells ◽  
R. K. O'Dor ◽  
S. K. Buckley
Keyword(s):  
Protein Synthesis ◽  
High Rate ◽  
Follicle Cells ◽  
Octopus Vulgaris ◽  
Amino Acid Incorporation ◽  
Gland Extract ◽  
Acid Incorporation ◽  
Kinetics Of

1. Protein synthesis occurs at a high rate in the ovaries of maturing Octopus vulgaris and can be measured from the incorporation of [14C]leucine in vivo and in isolated groups of eggs in vitro. 2. Removal of the optic glands in vivo 1--3 days prior to testing markedly reduces amino acid incorporation in vivo or in vitro. After 5 days in vivo incorporation stops. 3. The rate of incorporation in vitro is increased by the addition of optic gland extract. 4. Analysis of the kinetics of leucine uptake and incorporation in vitro indicates that the hormone has an effect on the inward transport of leucine which is independent of its action on protein synthesis. 5. Electron-microscope studies of the follicle cells and ova show that the former are the site of protein synthesis. 6. Changes in either uptake or incorporation into protein by the follicle cells can be used as a qualitative biolobical assay for the optic gland hormone. Uptake is very easy to measure but incorporation is the more sensitive parameter. Either is potentially suitable as a quantitative assay for this and perhaps also for other molluscan gonadotropins.

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