A model explaining the mechanism of initiation of differentiation is presented. It is based on the finding that sporulation in B. subtilis can be induced by the same nutrient deficiencies that also induce the synthesis of highly phosphorylated nucleotides. Two of these nucleotides are synthesized by membrane vesicles. Synthesis can be inhibited by the same metabolites of glucose that also inhibit sporulation. It is concluded, therefore, that the plasma membrane synthesizes unusual nucleotides in response to nutrient deficiencies. By several as yet unknown steps, these nucleotides then cuase changes in the metabolism of the organism leading to the formation of spores. Both structure and mechanism of synthesis of adenosine hexaphosphate, pppAppp, have been elucidated by use of ATP analogues.
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http://dx.doi.org/10.1073/pnas.73.2.331 | DOI Listing |
Nucleic Acids Res
January 2025
Department of Pharmaceutical Sciences, University of California, Irvine, CA 92697-3958, United States.
Growth in the development of engineered polymerases for synthetic biology has led to renewed interest in assays that can measure the fidelity of polymerases that are capable of synthesizing artificial genetic polymers (XNAs). Conventional approaches require purifying the XNA intermediate of a replication cycle (DNA → XNA → DNA) by denaturing polyacrylamide gel electrophoresis, which is a slow, costly, and inefficient process that requires a large-scale transcription reaction and careful extraction of the XNA strand from the gel slice. In an effort to streamline the assay, we developed a purification-free approach in which the XNA transcription and reverse transcription steps occur inside the matrix of a hydrogel-coated magnetic particle.
View Article and Find Full Text PDFBiochemistry (Mosc)
December 2024
National Research Centre "Kurchatov Institute", Moscow, 123182, Russia.
Bacterial and viral RNA polymerases are promising targets for the development of new transcription inhibitors. One of the potential blockers of RNA synthesis is 7,8-dihydro-8-oxo-1,-ethenoadenine (oxo-εA), a synthetic compound that combines two adenine modifications: 8-oxoadenine and 1,-ethenoadenine. In this study, we synthesized oxo-εA triphosphate (oxo-εATP) and showed that it could be incorporated by the RNA-dependent RNA polymerase of SARS-CoV-2 into synthesized RNA opposite template residues A and G in the presence of Mn ions.
View Article and Find Full Text PDFBiochemistry (Mosc)
December 2024
Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, 630090, Russia.
Taking into account involvement of the RNA-binding proteins in regulation of activity of poly(ADP-ribose) polymerase 1 (PARP1), a key factor of DNA repair, the effect of the intrinsically disordered protein Sam68 (Src-associated substrate during mitosis of 68 kDa) on catalytic activity of this enzyme was studied. Plasmid containing coding sequence of the Sam68 protein was obtained. Using the obtained construct, conditions for the Sam68 expression in cells were optimized and procedure for protein purification was developed.
View Article and Find Full Text PDFViruses
December 2024
School of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, China.
De novo synthesis of phage genomes enables flexible genome modification and simplification. This study explores the synthetic genome assembly of phage vB_PaeS_SCUT-S4 (S4), a 42,932 bp headful packaging phage, which encapsidates a terminally redundant, double-stranded DNA genome exceeding unit length. We demonstrate that using the yeast TAR approach, the S4 genome can be assembled and rebooted from a unit-length genome plus a minimal 60 bp terminal redundant sequence.
View Article and Find Full Text PDFNutrients
January 2025
University Centre for Prevention and Sports Medicine, Department of Orthopaedics, Balgrist University Hospital, University of Zurich, 8008 Zurich, Switzerland.
Background/objectives: In recent years, there has been a growing interest in precision nutrition and its potential for disease prevention. Differences in individual responses to diet, especially among populations of different ancestry, have underlined the importance of understanding the effects of genetic variations on nutrient intake (nutrigenomics). Since humans generally cannot synthesize essential vitamins, the maintenance of healthy bodily functions depends on dietary vitamin intake.
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