scholarly journals Microtubule protein synthesis during oogenesis and early embryogenesis in Xenopus laevis

1975 ◽  
Vol 145 (3) ◽  
pp. 527-534 ◽  
Author(s):  
R Q. W. Pestell
Keyword(s):  
Protein Synthesis ◽  
Xenopus Laevis ◽  
Soluble Protein ◽  
De Novo ◽  
Early Embryogenesis ◽  
Microtubule Polymerization ◽  
Microtubule Protein

A method is described which permits the preparation of descrete classes of oocytes of different sizes from all stages of oogenesis in Xenopus laevis. This technique is used in the determination of the content of microtubule protein in oocytes during the course of oogenesis. These experiments show that microtubule protein is present in oocytes of all sizes assayed and that the amount is simply related to the volume of the oocyte. In the largest oocytes microtubule protein constitutes 1% of the soluble protein and this amount does not change on maturation and fertilization. These results show that the changes occurring in the oocyte on maturation which allow the cytoplasm to support microtubule polymerization occur as a result of a modification of the pre-existing microtubule protein, not from protein synthesis de novo. These experiments also indicate that the synthesis of microtubule protein either form ‘masked’ mRNA or from newly synthesized mRNA plays an insignificant role in microtubule protein synthesis at maturation, ovulation and immediately post-fertilization.

Gene expression in Xenopus embryogenesis

Development ◽  
1985 ◽  
Vol 89 (Supplement) ◽  
pp. 113-124
Author(s):  
Igor B. Dawid ◽  
Susan R. Haynes ◽  
Milan Jamrich ◽  
Erzsebet Jonas ◽  
Seiji Miyatani ◽  
...  
Keyword(s):  
Gene Expression ◽  
Protein Synthesis ◽  
Xenopus Laevis ◽  
Cdna Cloning ◽  
Early Embryogenesis ◽  
Gene Activity ◽  
Complex Nature ◽  
Cdna Clones ◽  
Midblastula Transition ◽  
First Time

This article considers some aspects of the storage of macromolecules in the oocyte of Xenopus laevis and the activation of previously unexpressed genes during early embryogenesis. The large quantity and complex nature of poly(A)+ RNA accumulated in the egg provides the cleavage embryo with a supply of mRNA sufficient to sustain protein synthesis for several hours of development. Onset of gene activity at the midblastula transition (MBT) leads to the synthesis and accumulation of molecules of various RNA classes, including tRNAs, rRNAs, mRNAs and mitochondrial RNAs. At gastrulation the poly(A)+ RNA population is still qualitatively similar to that of the egg but some sequences not present in egg RNA have accumulated by this time. Through the use of a subtractive cDNA cloning procedure we have prepared a library of sequences that represent genes activated for the first time between MBT and gastrula. A study of several of these cDNA clones suggests that genes in this class are restricted in their activity to embryonic and tadpole stages.

Evidence for de novo synthesis of an aflatoxin pathway methyltransferase near the cessation of active growth and the onset of aflatoxin biosynthesis in Aspergillus parasiticus mycelia

1990 ◽  
Vol 36 (1) ◽  
pp. 1-5 ◽  
Author(s):  
T. E. Cleveland ◽  
D. Bhatnagar
Keyword(s):  
Protein Synthesis ◽  
Soluble Protein ◽  
Aspergillus Parasiticus ◽  
De Novo ◽  
Protein Fractions ◽  
Time Interval ◽  
Aflatoxin Biosynthesis ◽  
Active Growth ◽  
Mtase Activity ◽  
De Novo Protein Synthesis

The accumulation of both activity and protein of a methyltransferase (MTase) from Aspergillus parasiticus, which catalyzes conversion of sterigmatocystin to O-methylsterigmatocystin in the aflatoxin pathway, was detected in fungal mycelia slightly before the onset of aflatoxin biosynthesis in the same cultures. MTase protein was identified in mycelial postmicrosomal (soluble protein) fractions by electrophoresis and subsequent immunoblotting using antiserum raised against purified MTase protein; MTase activity was determined by measuring the rate of conversion of sterigmatocystin to O-methylsterigmatocystin in the presence of soluble protein fractions. Using the above technique, it was determined that MTase protein as well as MTase activity increased sharply in mycelia 30 to 45 h after inoculation, shortly after which, mycelial growth rate began to decline. During the subsequent time interval (45 to 70 h after inoculation), a sharp increase in aflatoxin levels was detected in the culture medium. Results obtained from an experiment in which cycloheximide was added to cultures at various times to inhibit protein synthesis and from an experiment in which mycelial proteins were radiolabelled to identify newly synthesized proteins indicated that accumulation of MTase activity and protein in late growth phase mycelia is due to de novo protein synthesis. Key words: aflatoxin, methyltransferase, biosynthetic pathway.

Synthesis, storage, and release of MSH in the pars intermedia of the pituitary gland of Xenopus laevis during background adaptation

1977 ◽  
Vol 55 (6) ◽  
pp. 922-927 ◽  
Author(s):  
B. G. Jenks ◽  
A. P. VanOverbeeke ◽  
B. F. McStay
Keyword(s):  
Protein Synthesis ◽  
Xenopus Laevis ◽  
De Novo ◽  
Synthetic Activity ◽  
White Background ◽  
Pars Intermedia ◽  
Black Background ◽  
Background Adaptation ◽  
Initial Stage ◽  
Simultaneous Decrease

Pituitary levels of melanophore-stimulating hormone (MSH), release of MSH, and protein synthetic activity in the pars intermedia were determined in Xenopus laevis during background adaptation. MSH was measured using a radioimmunoassay to α-MSH; uptake of [3H]lysine, determined autoradiographically, was used to assess protein synthesis; changes in melanophore index indicated changes in release of MSH. Adaptation to black background led to eventual depletion of MSH and increased protein synthetic activity. Conversely, during adaptation to a white background MSH levels increased and protein synthesis decreased. Changes in synthesis lagged considerably behind changes in release. During the initial stage of black-background adaptation, release of MSH was not accompanied by simultaneous decrease in levels of MSH in the gland from which it is concluded that replenishment of MSH took place. Our results indicated that this replenishment in the gland could not be accounted for by de novo synthesis of the hormone. It is proposed that a stored precursor to MSH exists, conversion of which provides for rapid replenishment of MSH. It is suggested that the factor(s) controlling MSH release affect synthesis of this hormone only indirectly.

Preparation, spectral properties and biological activities of 5-bromo-6-methyl-2'-deoxyuridine and 5-iodo-6-methyl-2'-deoxyuridine

1980 ◽  
Vol 45 (8) ◽  
pp. 2364-2370 ◽  
Author(s):  
Antonín Holý ◽  
Erik De Clercq
Keyword(s):  
Antiviral Activity ◽  
Spectral Properties ◽  
De Novo ◽  
Biological Activities ◽  
Rabbit Kidney ◽  
Kidney Cells ◽  
Cd Spectra ◽  
Methyl Derivatives ◽  
Primary Rabbit

Reaction of 3',5'-di-O-benzoyl-6-methyl-2'-deoxyuridine (IIa) with elementary bromine or iodine afforded 5-halogeno derivatives IIc and IId which on methanolysis gave 5-bromo-6-methyl-2'-deoxyurine (Ic) and 5-iodo-6-methyl-2'-deoxyurine (Id), respectively. The CD spectra of Ic, Id and 6-methyl-2'-deoxyuridine (Ia) are compared and discussed with regard to determination of the nucleoside conformation. Unlike 5-bromo- and 5-iodo-2'-deoxyuridine, the 6-methyl derivatives Ic and Id exhibit neither antibacterial nor antiviral activity. Nor do they exert any antimetabolic effect on the de novo DNA synthesis in primary rabbit kidney cells.

In vivo study of the biosynthesis of long-chain fatty acids using deuterated water

1995 ◽  
Vol 269 (2) ◽  
pp. E247-E252 ◽  
Author(s):  
H. O. Ajie ◽  
M. J. Connor ◽  
W. N. Lee ◽  
S. Bassilian ◽  
E. A. Bergner ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
De Novo ◽  
Chain Elongation ◽  
Deuterated Water ◽  
De Novo Synthesis ◽  
Isotopomer Distribution ◽  
Long Chain ◽  
Mass Isotopomer

To determine the contributions of preexisting fatty acid, de novo synthesis, and chain elongation in long-chain fatty acid (LCFA) synthesis, the synthesis of LCFAs, palmitate (16:0), stearate (18:0), arachidate (20:0), behenate (22:0), and lignocerate (24:0), in the epidermis, liver, and spinal cord was determined using deuterated water and mass isotopomer distribution analysis in hairless mice and Sprague-Dawley rats. Animals were given 4% deuterated water for 5 days or 8 wk in their drinking water. Blood was withdrawn at the end of these times for the determination of deuterium enrichment, and the animals were killed to isolate the various tissues for lipid extraction for the determination of the mass isotopomer distributions. The mass isotopomer distributions in LCFA were incompatible with synthesis from a single pool of primer. The synthesis of palmitate, stearate, arachidate, behenate, and lignocerate followed the expected biochemical pathways for the synthesis of LCFAs. On average, three deuterium atoms were incorporated for every addition of an acetyl unit. The isotopomer distribution resulting from chain elongation and de novo synthesis can be described by the linear combination of two binomial distributions. The proportions of preexisting, chain elongation, and de novo-synthesized fatty acids as a percentage of the total fatty acids were determined using multiple linear regression analysis. Fractional synthesis was found to vary, depending on the tissue type and the fatty acid, from 47 to 87%. A substantial fraction (24-40%) of the newly synthesized molecules was derived from chain elongation of unlabeled (recycled) palmitate.

scholarly journals nanotatoR: a tool for enhanced annotation of genomic structural variants

BMC Genomics ◽  
2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Surajit Bhattacharya ◽  
Hayk Barseghyan ◽  
Emmanuèle C. Délot ◽  
Eric Vilain
Keyword(s):  
De Novo ◽  
Genome Mapping ◽  
Gene List ◽  
Sufficient Information ◽  
Rna Seq ◽  
Structural Variants ◽  
De Novo Genome Assembly ◽  
Pathogenic Variants ◽  
Increased Sensitivity

Abstract Background Whole genome sequencing is effective at identification of small variants, but because it is based on short reads, assessment of structural variants (SVs) is limited. The advent of Optical Genome Mapping (OGM), which utilizes long fluorescently labeled DNA molecules for de novo genome assembly and SV calling, has allowed for increased sensitivity and specificity in SV detection. However, compared to small variant annotation tools, OGM-based SV annotation software has seen little development, and currently available SV annotation tools do not provide sufficient information for determination of variant pathogenicity. Results We developed an R-based package, nanotatoR, which provides comprehensive annotation as a tool for SV classification. nanotatoR uses both external (DGV; DECIPHER; Bionano Genomics BNDB) and internal (user-defined) databases to estimate SV frequency. Human genome reference GRCh37/38-based BED files are used to annotate SVs with overlapping, upstream, and downstream genes. Overlap percentages and distances for nearest genes are calculated and can be used for filtration. A primary gene list is extracted from public databases based on the patient’s phenotype and used to filter genes overlapping SVs, providing the analyst with an easy way to prioritize variants. If available, expression of overlapping or nearby genes of interest is extracted (e.g. from an RNA-Seq dataset, allowing the user to assess the effects of SVs on the transcriptome). Most quality-control filtration parameters are customizable by the user. The output is given in an Excel file format, subdivided into multiple sheets based on SV type and inheritance pattern (INDELs, inversions, translocations, de novo, etc.). nanotatoR passed all quality and run time criteria of Bioconductor, where it was accepted in the April 2019 release. We evaluated nanotatoR’s annotation capabilities using publicly available reference datasets: the singleton sample NA12878, mapped with two types of enzyme labeling, and the NA24143 trio. nanotatoR was also able to accurately filter the known pathogenic variants in a cohort of patients with Duchenne Muscular Dystrophy for which we had previously demonstrated the diagnostic ability of OGM. Conclusions The extensive annotation enables users to rapidly identify potential pathogenic SVs, a critical step toward use of OGM in the clinical setting.

scholarly journals De novo expression and antibacterial potential of four lactoferricin peptides in cell-free protein synthesis system

2021 ◽  
Vol 29 ◽  
pp. e00583
Author(s):  
Nawal Abd El-Baky ◽  
Maie Ahmed Elkhawaga ◽  
Eman Shawky Abdelkhalek ◽  
Mona Mohammed Sharaf ◽  
Elrashdy Mustafa Redwan ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
De Novo ◽  
Synthesis System ◽  
Free Protein ◽  
Protein Synthesis System ◽  
Cell Free Protein Synthesis ◽  
Antibacterial Potential
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