Stimulation of globin synthesis by RNA purified from reticulocyte supernatant fraction

doi:10.1016/0003-9861(68)90616-4

Archives of Biochemistry and Biophysics

Volume 125, Issue 2, May 1968, Pages 580-593

To Richard S. Schweet, pioneer in the field of peptide synthesis—in memoriam.

https://doi.org/10.1016/0003-9861(68)90616-4 Get rights and content

Abstract

Rabbit reticulocytes were preincubated in sodium fluoride to dissociate polyribosomes to 80S ribosomes, and RNA was phenol-extracted from the supernatant fraction of these cells after removing ribosomes. This purified RNA was fractionated by sucrose density gradient centrifugation, and found to contain a fraction corresponding to 6S to 14S which induced formation of polyribosomes active in protein synthesis and specifically stimulated rabbit α- and β-globin chain formation at least 2.5-fold in a cell-free system containing ribosomes prepared from fluoride-treated rabbit reticulocytes. No stimulation of protein synthesis was observed with addition of the RNA fraction to a cell-free system containing ribosomes prepared from reticulocytes not preincubated with sodium fluoride. Addition of this rabbit fluoride RNA to a mouse cell-free system stimulated amino acid incorporation into protein, but no formation of rabbit globin chains was detectable.

References (44)

J. Bishop et al.
Nature
(1961)
J. Kruh et al.
Biochim. Biophys. Acta
(1961)
H. Lamfrom
J. Mol. Biol
(1961)
H.R.V. Arnstein et al.
Nature
(1962)
J. Bishop et al.
Biochim. Biophys. Acta
(1962)
J. Kruh et al.
Biochim. Biophys. Acta
(1962)
A.J. Munro et al.
Nature
(1963)
H.R.V. Arnstein et al.
Biochem. J
(1964)
J. Kruh et al.
Biochim. Biopys. Acta
(1964)
J. Kruh et al.
Biochim. Biophys. Acta
(1964)

L. Luzzatto et al.

J. Schaeffer et al.

Biochim. Biophys. Acta

(1964)

G. Brawerman et al.

Biochim. Biophys. Acta

(1965)

G. Marbaix et al.

Biochim. Biophys. Acta

(1966)

G. Schapira et al.

J. Mol. Biol

(1966)

A.S. Weisberger et al.

L. Grossbard et al.

J.C. Drach et al.

Biochim. Biophys. Acta

(1964)

J.A. Hunt et al.

Biochemistry

(1967)

P.A. Marks et al.

H. Fraenkel-Conrat et al.

Virology

(1961)

H. Borsook et al.

J. Biol. Chem

(1957)

Cited by (6)

Presence of haemoglobin messenger RNA in the postribosomal supernatant of rabbit reticulocytes and conditions necessary for its translation
1972, FEBS Letters
RNA synthesized during differentiation of yolk sac erythroid cells
1971, Journal of Molecular Biology
Yolk sac erythroid cells from fetal mice at 11 days gestation (total gestation period is 20 to 2 days) synthesize proteins, approximately two-thirds of which are non-hemoglobin proteins and the remainder hemoglobins. Between day 11 and day 13, there is a decrease in the rate of synthesis of all types of RNA, with the decline in the rate of formation of 45 s RNA being greater than that of heterogeneously sedimenting RNA. In 11-day cells, at least two species of RNA are formed with an electrophoretic mobility between 18 s and 5 s RNA, corresponding to 14 s and 9 s RNA. The 14 s and 9 s RNA's are associated with polyribosomes and are shown not to be degradation products of ribosomal RNA. As the yolk sac erythroid cells differentiate, between days 11 and 13, the rate of synthesis of 9 s RNA declines markedly. On day 13, non-hemoglobin protein synthesis has declined to nil, while the rate of synthesis of hemoglobins is essentially unchanged from that of day 11. In 11-day yolk sac erythroid cells incubated with actinomycin D or with hydroxyurea for 60 minutes, newly synthesized 9 s RNA disappears completely, whereas the newly formed 18 s or 14 s RNA is relatively stable. Concomitantly, actinomycin D or hydroxyurea causes a marked inhibition of formation of nuclear proteins, but not of cytoplasmic protein (hemoglobins) synthesis. It is suggested that the 9 s RNA contains messenger RNA for nuclear proteins.
Ribonuclease activity in the reticulocyte cell-free system
1970, BBA Section Nucleic Acids And Protein Synthesis
Ribonuclease activity in the reticulocyte cell-free system is operative during the period when protein synthesis occurs. The ribonuclease in the system is not active at 0°, does not require energy, and degrades rRNA at a rate greater than soluble RNA. Supplementation of the reticulocyte lysate system with reticulocyte RNA stimulated protein synthesis and simultaneously inhibited the inherent ribonuclease activity of the system. Non-reticulocyte RNA also inhibited ribonuclease activity in this system, as did the nonpolynucleotides hemin and Co²⁺.
Globin α-Chain synthesis directed by “supernatant” 10S RNA from rabbit reticulocytes
1972, Nature New Biology
Identification of mouse haemoglobin messenger RNA
1971, Nature New Biology
Translation of rabbit haemoglobin messenger RNA by thalassaemic and non-thalassaemic ribosomes
1971, Nature New Biology

^☆: These studies were supported, in part by grants from the U.S. Public Health Service (GM-14552-02) and the National Science Foundation (GB-4631).

²: Trainee in Hematology, supported by a grant from N.I.A.M.D. (TIAM-5231-08).

View full text

Stimulation of globin synthesis by RNA purified from reticulocyte supernatant fraction☆

Abstract

Nature

Biochim. Biophys. Acta

J. Mol. Biol

Nature

Biochim. Biophys. Acta

Biochim. Biophys. Acta

Nature

Biochem. J

Biochim. Biopys. Acta

Biochim. Biophys. Acta

Biochim. Biophys. Acta

Biochim. Biophys. Acta

Biochim. Biophys. Acta

J. Mol. Biol

Biochim. Biophys. Acta

Biochemistry

Virology

J. Biol. Chem