Small peptides bound to polysomal RNA inhibit gene expression in cell-free systems, replication of stimulated lymphocytes and DNA repair in isolated chromatin.
Polysomal poly(A)+-RNA prepared from isolated calf liver polysomes by deproteinization and affinity chromatography on oligo(dT)-Sepharose at pH 6 contains low molecular weight peptides (between 600-1500 daltons) bound noncovalently. These peptides were extracted from the poly(A)+-RNA-peptides complex by precipitation of the nucleic acids with 80% (v/v) ethanol at alkaline pH (9.5) and purified on Sephadex G-25 and G-15 columns. Further fractionation was performed by silica gel chromatography and high performance liquid chromatography (h.p.l.c.). The amino acid composition of the isolated peptidic fraction was compared with similar peptides obtained from rat liver, rabbit reticulocyte and calf thymus polysomes. Effluent (ribosomal) RNA contains only negligible amount of peptides. Isolated polysomal RNA peptides were named "deprimerones" (from Latin "deprimere"), since they have a general depressing effect on gene expression in vitro (Hillar & Przyjemski, 1979). Isolated deprimerones not only inhibit DNA transcription, RNA translation in reconstituted cell-free systems, but also DNA replication by DNA polymerase beta with single- and double-stranded DNA template and synthetic deoxyribonucleotide polymers. The inhibitory effect on replication was correlated with the inhibition of [3H]-deoxyribonucleotide incorporation in isolated chromatin and in stimulated lymphocyte cell cultures. The isolated deprimerones are characterized by similar amino acid compositions in various species.
Hillar
M
M
Stolzman
Z
Z
Santarelli
I
I
Patt
L M
LM
Houck
J C
JC
Chan
J Y
JY
Wyborny
L E
LE
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002417:Cattle; D002455:Cell Division; D002474:Cell-Free System; D002843:Chromatin / Q000378:metabolism* / Q000648:ultrastructure; D004260:DNA Repair; D008099:Liver / Q000378:metabolism; D008214:Lymphocytes / Q000166:cytology / Q000378:metabolism*; D010455:Peptides / Q000302:isolation & purification / Q000502:physiology*; D011061:Poly A / Q000235:genetics* / Q000378:metabolism; D011132:Polyribosomes / Q000378:metabolism*; D012313:RNA / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger; D011817:Rabbits; D051381:Rats; D012156:Reticulocytes / Q000378:metabolism; D013045:Species Specificity; D013950:Thymus Gland / Q000378:metabolism; D014158:Transcription, Genetic
D000596:Amino Acids; D002843:Chromatin; D010455:Peptides; D012333:RNA, Messenger; D011061:Poly A; D012313:RNA