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P M Prichard

Publications and source records attributed to P M Prichard.

12 recordsLinked to original sources
PubMed1975

Increased translation as a result of elevated initation factor activity after administration of 3-methylcholanthrene.

Protein synthesis initiation factors present in a crude 0.5 M KCL microsomal wash fraction were isolated from the livers of immature rats that had been injected either 2 or 17 hours earlier with the plycyclic hydrocarbon, 3-methylcholanthrene (3MC), and then were tested for their ability to stimulate natural mRNA-directed protein synthesis in vitro. After purification of the initiation factors by means of ammonium sulfate fractionation, and DEAE-cellulose and Sephadex G-200 chromatography, M2A and M2B, but not M3 or M1, from the livers of 3MC-pretreated rats were more active than were similarly-prepared control factors in their ability to initiate the synthesis of rabbit globin polypeptides in a highly fractionated cell-free protein synthesizing system derived from rabbit reticulocytes. The greater activity of the M2A and M2B preparations from drug-treated rat liver did not appear to be due to differences in the composition of initiation factor protein extracted from the livers of control or experimental rats and occurred very early after administration of 3MC, i.e., 2 hr. The role of the protein synthesis initiation factors in the altered rates of protein synthesis which accompany cytodifferentiation and growth is discussed.

Ammonium Sulfate↗
PubMed1972

Separation of reticulocyte initiation factor M 2 activity into two components.

IF-M(2), one of three initiation factors isolated by DEAE-cellulose chromatography from the 0.5 M KCl-wash fraction of rabbit reticulocyte ribosomes, has been separated by Sephadex G-200 chromatography into two components: IF-M(2A) and IF-M(2B). IF-M(2A) elutes near the void-volume, while IF-M(2B), which is much smaller in molecular weight than IF-M(2A), elutes slightly after a hemoglobin marker. In the presence of the other appropriate factors, both IF-M(2A) and IF-M(2B) are required to stimulate poly(U)-directed polyphenylalanine synthesis at low Mg(++) concentration, ApUpG-directed Met-tRNA(F) binding to washed reticulocyte ribosomes, and initiation of globin synthesis from endogenous mRNA. IF-M(2A) stimulates ribosome-dependent GTP hydrolysis, while IF-M(2B) does not; IF-M(2B) stimulates ApUpG-directed fMet-tRNA(F) binding in the presence of IF-M(1), while IF-M(2A) does not. Although IF-M(2A) and IF-M(2B) can be distinguished from each other by size and by activity, a distinct function for IF-M(2B) has not yet been found. Therefore, its precise role in the initiation process remains unclear.

Animals↗
PubMed1971

Translation of exogenous messenger RNA for hemoglobin on reticulocyte and liver ribosomes.

The ribosome and initiation factor requirements for translation of rabbit-reticulocyte hemoglobin mRNA on rabbit reticulocyte ribosomes, reticulocyte ribosomal subunits, and liver ribosomes have been studied. Excellent synthesis of globin chains from exogenous mRNA in the fractionated cell-free system has been achieved. There is a near absolute requirement for each of the initiation factors, M(1), M(2), and M(3) (as well as for the supernatant proteins) for the translation of exogenous mRNA. Liver microsomal wash will partially replace reticulocyte factors M(1) and M(2), but will not replace the requirement for reticulocyte factor M(3). Rabbit liver ribosomes and rabbit reticulocyte ribosomes are equally active in their ability to support the translation of exogenous hemoglobin mRNA.

Animals↗
PubMed1971

Initial dipeptide formation in hemoglobin biosynthesis.

Initiation factors M(1) + M(2) from reticulocyte ribosomes bind Met-tRNA(F) to rabbit reticulocyte ribosomes containing endogenous hemoglobin mRNA. The initial binding of Met-tRNA(F) appears to be to the small ribosomal subunit. The Met-tRNA(F) is able to participate in what is presumed to be the first peptide bond in the formation of hemoglobin, namely the synthesis of a methionyl-valine dipeptide. The formation of this methionyl-valine dipeptide requires Met-tRNA(F), initiation factors M(1), M(2), and M(3), as well as Val-tRNA and T(1). No synthesis of methionyl-valine dipeptide takes place if Met-tRNA(F) is replaced by Met-tRNA(M), or if initiation factor M(3) is omitted. Thus, Met-tRNA(F) appears to be the initiator tRNA for hemoglobin biosynthesis and M(3), although required for the synthesis of the first peptide bond of hemoglobin, does not appear to be necessary, under the experimental conditions studied, for Met-tRNA(F) binding.

Animals↗