Total poly(A)+ RNA was isolated from B cell hybridomas producing monoclonal antibodies of the IgM class specific for sheep red blood cells. These RNAs were microinjected into Xenopus oocytes and the translation products were analyzed. Unlike the hybridoma cells, the oocytes synthetized only the light chains and oligomer IgM, but evidence for assembly of functional pentamer IgM was not obtained. The possible reasons for these differences are discussed.

• MeSH
• B-Lymphocytes immunology   MeSH
• Hybridomas immunology   MeSH
• Immunoglobulin G biosynthesis   MeSH
• Immunoglobulin M biosynthesis   genetics   MeSH
• RNA, Messenger genetics   MeSH
• Oocytes metabolism   MeSH
• Sheep MeSH
• Poly A genetics   MeSH
• Protein Biosynthesis MeSH
• RNA genetics   MeSH
• Xenopus MeSH
• Animals MeSH
• Check Tag
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Comparative Study MeSH
• Names of Substances
• Immunoglobulin G MeSH
• Immunoglobulin M MeSH
• RNA, Messenger MeSH
• Poly A MeSH
• RNA MeSH

RNA synthesis in 2-32 cell embryos, as assessed by alpha-amanitin-sensitive 3H-uridine incorporation, was first detectable in 4-cell stage rabbit and 8-cell stage sheep embryos. In the rabbit, uridine incorporation was detectable at the 2-cell stage but was unaltered by alpha-amanitin, indicating synthesis of non-polymerase II-dependent RNA species. Initiation of mRNA synthesis as determined by in situ hybridization with 3H-poly (U) probe was first detectable in late 2-cell stage rabbit and 4-cell stage sheep embryos. In the rabbit embryos, nuclear labelling increased from the late 2-cell stage to the 16-cell stage, following a pattern similar to that of 3H-uridine incorporation. In contrast, the intensity of cytoplasmic labelling decreased from the 1- to the 8-cell stage and then increased up to the 32-cell stage. In sheep embryos, nuclear labelling by the poly (U) probe increased from the 4- to the 16-cell stage. It is concluded that initiation of transcription of the embryonic genome (mRNA) can be detected via the current methods used at the 4-cell stage in the rabbit and the 8-cell stage in the sheep.

• MeSH
• Amanitins pharmacology   MeSH
• Autoradiography MeSH
• Blastocyst metabolism   MeSH
• Species Specificity MeSH
• Transcription, Genetic drug effects   MeSH
• In Situ Hybridization MeSH
• Rabbits embryology   metabolism   MeSH
• RNA, Messenger metabolism   MeSH
• Sheep embryology   metabolism   MeSH
• Poly A metabolism   MeSH
• RNA biosynthesis   MeSH
• Animals MeSH
• Check Tag
• Rabbits embryology   metabolism   MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Comparative Study MeSH
• Names of Substances
• Amanitins MeSH
• RNA, Messenger MeSH
• Poly A MeSH
• RNA MeSH

• MeSH
• Carbodiimides metabolism   MeSH
• Magnetics MeSH
• RNA, Messenger isolation & purification   MeSH
• Myocardium chemistry   MeSH
• Blotting, Northern MeSH
• Oligodeoxyribonucleotides metabolism   MeSH
• Poly A chemistry   metabolism   MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Carbodiimides MeSH
• RNA, Messenger MeSH
• oligo (dT) MeSH Browser
• Oligodeoxyribonucleotides MeSH
• Poly A MeSH

Polyoma (Py) virus-specific RNA, synthesized at reduced level in infected cells in the presence of antiviral compound 5-bromo-2'-deoxyuridine (BrdUrd) was characterized in more detail by Northern blot hybridization analysis. The results obtained with total, cytoplasmic and poly(A)RNA, isolated from mouse embryo cell cultures 42 hrs p.i. indicate that BrdUrd (6.34 micrograms/ml) lowers the level of typical classes of major virus DNA transcripts to a similar extent and that no new, atypical transcription products are formed.

• MeSH
• Bromodeoxyuridine pharmacology   MeSH
• Transcription, Genetic drug effects   MeSH
• RNA, Messenger biosynthesis   MeSH
• Blotting, Northern MeSH
• Poly A biosynthesis   MeSH
• Polyomavirus drug effects   genetics   metabolism   MeSH
• Gene Expression Regulation, Viral drug effects   MeSH
• Virus Replication drug effects   MeSH
• RNA, Viral biosynthesis   MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Bromodeoxyuridine MeSH
• RNA, Messenger MeSH
• Poly A MeSH
• RNA, Viral MeSH

Poly(A) RNA was isolated from the gastric mucosa of the bovine fourth stomach (the abomasum) using and analysing several calves not older than 12 days. The amount of the preprochymosin mRNA in the mucosa of those animals at best reaches about 5-10% of the poly(A) RNA as estimated by in vitro translation and immunoprecipitation. Starting from that material double-stranded complementary DNA was synthesized, inserted by dG dC tailing into the PstI site of the vector plasmid pBR322 and used for transformation of E. coli. Tetracycline resistant clones containing DNA sequences coding for the full length of prochymosin were recognized by colony hybridization with five specific d-oligonucleotides corresponding either to the N-terminal, the middle or the C-terminal part of prochymosin. Six recombinants were detected by screening of 1 500 recombinants with an oligonucleotide which corresponds to positions 649 to 663 of the nucleotide sequence published by Harris et al. (1982). Two of them were found to cover together the complete prochymosin sequence as evidenced by both positive colony hybridization with either the N-terminal or the C-terminal oligonucleotide probe, as well as by the restriction pattern of the selected plasmids.

• MeSH
• Chymosin analysis   genetics   MeSH
• DNA genetics   MeSH
• Escherichia coli genetics   MeSH
• Cloning, Molecular MeSH
• RNA, Messenger genetics   isolation & purification   MeSH
• Oligonucleotides analysis   MeSH
• Plasmids MeSH
• Poly A genetics   isolation & purification   MeSH
• Enzyme Precursors genetics   MeSH
• DNA Restriction Enzymes MeSH
• Base Sequence MeSH
• Cattle genetics   metabolism   MeSH
• Abomasum analysis   MeSH
• Gastric Mucosa analysis   MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Cattle genetics   metabolism   MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Chymosin MeSH
• DNA MeSH
• RNA, Messenger MeSH
• Oligonucleotides MeSH
• Poly A MeSH
• Enzyme Precursors MeSH
• preprorennin MeSH Browser
• prorennin MeSH Browser
• DNA Restriction Enzymes MeSH

BACKGROUND: Limited accessibility to intestinal epithelial tissue in wild animals and humans makes it challenging to study patterns of intestinal gene regulation, and hence to monitor physiological status and health in field conditions. To explore solutions to this limitation, we have used a noninvasive approach via fecal RNA-seq, for the quantification of gene expression markers in gastrointestinal cells of free-range primates and a forager human population. Thus, a combination of poly(A) mRNA enrichment and rRNA depletion methods was used in tandem with RNA-seq to quantify and compare gastrointestinal gene expression patterns in fecal samples of wild Gorilla gorilla gorilla (n = 9) and BaAka hunter-gatherers (n = 10) from The Dzanga Sangha Protected Areas, Central African Republic. RESULTS: Although only a small fraction (< 4.9%) of intestinal mRNA signals was recovered, the data was sufficient to detect significant functional differences between gorillas and humans, at the gene and pathway levels. These intestinal gene expression differences were specifically associated with metabolic and immune functions. Additionally, non-host RNA-seq reads were used to gain preliminary insights on the subjects' dietary habits, intestinal microbiomes, and infection prevalence, via identification of fungi, nematode, arthropod and plant RNA. CONCLUSIONS: Overall, the results suggest that fecal RNA-seq, targeting gastrointestinal epithelial cells can be used to evaluate primate intestinal physiology and gut gene regulation, in samples obtained in challenging conditions in situ. The approach used herein may be useful to obtain information on primate intestinal health, while revealing preliminary insights into foraging ecology, microbiome, and diet.

• Keywords
• Gene expression, Nonhuman primate, Noninvasive method, RNA-seq,
• MeSH
• Feces * MeSH
• Gastrointestinal Tract metabolism   MeSH
• Gorilla gorilla genetics   MeSH
• Humans MeSH
• RNA, Messenger genetics   MeSH
• Poly A genetics   MeSH
• RNA-Seq * MeSH
• Gene Expression Profiling * MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• RNA, Messenger MeSH
• Poly A MeSH

OTUD1 is a deubiquitinating enzyme involved in many cellular processes including cancer and innate, immune signaling pathways. Here, we perform a proximity labeling-based interactome study that identifies OTUD1 largely present in the translation and RNA metabolism protein complexes. Biochemical analysis validates OTUD1 association with ribosome subunits, elongation factors and the E3 ubiquitin ligase ZNF598 but not with the translation initiation machinery. OTUD1 catalytic activity suppresses polyA triggered ribosome stalling through inhibition of ZNF598-mediated RPS10 ubiquitination and stimulates formation of polysomes. Finally, analysis of gene expression suggests that OTUD1 regulates the stability of rare codon rich mRNAs by antagonizing ZNF598.

• Keywords
• ribosome stalling, translation, ubiquitination,
• MeSH
• Deubiquitinating Enzymes genetics   metabolism   MeSH
• Codon MeSH
• RNA, Messenger genetics   metabolism   MeSH
• Poly A * metabolism   MeSH
• Protein Biosynthesis MeSH
• Carrier Proteins * metabolism   MeSH
• Ubiquitination MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Deubiquitinating Enzymes MeSH
• Codon MeSH
• RNA, Messenger MeSH
• Poly A * MeSH
• Carrier Proteins * MeSH

• MeSH
• Humans MeSH
• RNA, Messenger MeSH
• Oncogenic Viruses * MeSH
• RNA, Neoplasm MeSH
• RNA, Viral MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• RNA, Messenger MeSH
• RNA, Neoplasm MeSH
• RNA, Viral MeSH

The 60 S viral RNA complex isolated from leukaemic plasma of chicken infected by avian myeloblastosis virus (AMV) was denatured, the poly(A)-RNA selected and centrifuged in a linear sucrose density gradient. RNA from each fraction was translated in vitro and the products were analyzed by slab polyacrylamide gel electrophoresis (PAGE). Unprocessed primary translation product (p64env) of MAV env gene from 21 S RNA fraction was immunoprecipitated by anti-gp85 serum. If, however, this RNA was translated in the presence of dog pancreas microsomal membranes (DPM), the processed 92 K MAV glycoprotein precursor (p92env) was immunoprecipitated by anti-gp85 serum. This precursor, unlike p64env was resistant to exogenous protease.

• MeSH
• Cell-Free System MeSH
• Gene Products, env biosynthesis   genetics   metabolism   MeSH
• Genes, env MeSH
• Glycosylation MeSH
• Chickens MeSH
• RNA, Messenger MeSH
• Poly A genetics   MeSH
• Protein Processing, Post-Translational * MeSH
• RNA Precursors blood   genetics   MeSH
• Protein Precursors biosynthesis   genetics   metabolism   MeSH
• Protein Biosynthesis * MeSH
• Avian Leukosis microbiology   MeSH
• Retroviridae Proteins biosynthesis   genetics   metabolism   MeSH
• Retroviridae Infections microbiology   MeSH
• RNA, Viral blood   genetics   MeSH
• RNA genetics   MeSH
• Avian Myeloblastosis Virus genetics   MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Gene Products, env MeSH
• RNA, Messenger MeSH
• Poly A MeSH
• RNA Precursors MeSH
• Protein Precursors MeSH
• Retroviridae Proteins MeSH
• RNA, Viral MeSH
• RNA MeSH

Regulation of mRNA translation by cytoplasmic polyadenylation is known to be important for oocyte maturation and further development. This process is generally controlled by phosphorylation of cytoplasmic polyadenylation element binding protein 1 (CPEB1). The aim of this study is to determine the role of Aurora kinase A in CPEB1 phosphorylation and the consequent CPEB1-dependent polyadenylation of maternal mRNAs during mammalian oocyte meiosis. For this purpose, we specifically inhibited Aurora kinase A with MLN8237 during meiotic maturation of porcine oocytes. Using poly(A)-test PCR method, we monitored the effect of Aurora kinase A inhibition on poly(A)-tail extension of long and short cyclin B1 encoding mRNAs as markers of CPEB1-dependent cytoplasmic polyadenylation. Our results show that inhibition of Aurora kinase A activity impairs neither cyclin B1 mRNA polyadenylation nor its translation and that Aurora kinase A is unlikely to be involved in CPEB1 activating phosphorylation.

• MeSH
• Aurora Kinase A metabolism   MeSH
• Cyclin B1 genetics   MeSH
• mRNA Cleavage and Polyadenylation Factors chemistry   metabolism   MeSH
• Phosphorylation MeSH
• Meiosis * MeSH
• RNA, Messenger metabolism   MeSH
• Oocytes enzymology   metabolism   MeSH
• Polyadenylation MeSH
• Sus scrofa metabolism   MeSH
• Animals MeSH
• Check Tag
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Aurora Kinase A MeSH
• Cyclin B1 MeSH
• mRNA Cleavage and Polyadenylation Factors MeSH
• RNA, Messenger MeSH