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961.
Ribonucleoside triphosphate (rNTP) incorporation in DNA by DNA polymerases is a frequent phenomenon that results in DNA structural change and genome instability. However, it is unclear whether the rNTP incorporation into DNA follows any specific sequence patterns. We analyzed multiple datasets of ribonucleoside monophosphates (rNMPs) embedded in DNA, generated from three rNMP-sequencing techniques. These rNMP libraries were obtained from Saccharomyces cerevisiae cells expressing wild-type or mutant replicative DNA polymerase and ribonuclease H2 genes. We performed computational analyses of rNMP sites around early and late-firing autonomously replicating sequences (ARSs) of the yeast genome, where leading and lagging DNA synthesis starts bidirectionally. We found the preference of rNTP incorporation on the leading strand in wild-type DNA polymerase yeast cells. The leading/lagging-strand ratio of rNTP incorporation changes dramatically within the first 1,000 nucleotides from ARSs, highlighting the Pol δ - Pol ϵ handoff during early leading-strand synthesis. Furthermore, the pattern of rNTP incorporation is markedly distinct between the leading and lagging strands not only in mutant but also in wild-type polymerase cells. Such specific signatures of Pol δ and Pol ϵ provide a new approach to track the labor of these polymerases. 相似文献
962.
Qian Wang Jin Liu Josephine M Janssen Francesca Tasca Hailiang Mei Manuel A F V Gonalves 《Nucleic acids research》2021,49(20):11986
Prime editing is a recent precision genome editing modality whose versatility offers the prospect for a wide range of applications, including the development of targeted genetic therapies. Yet, an outstanding bottleneck for its optimization and use concerns the difficulty in delivering large prime editing complexes into cells. Here, we demonstrate that packaging prime editing constructs in adenoviral capsids overcomes this constrain resulting in robust genome editing in both transformed and non-transformed human cells with up to 90% efficiencies. Using this cell cycle-independent delivery platform, we found a direct correlation between prime editing activity and cellular replication and disclose that the proportions between accurate prime editing events and unwanted byproducts can be influenced by the target-cell context. Hence, adenovector particles permit the efficacious delivery and testing of prime editing reagents in human cells independently of their transformation and replication statuses. The herein integrated gene delivery and gene editing technologies are expected to aid investigating the potential and limitations of prime editing in numerous experimental settings and, eventually, in ex vivo or in vivo therapeutic contexts. 相似文献
963.
Andrea Sacconi Claudia De Vitis Luisa de Latouliere Simona di Martino Francesca De Nicola Frauke Goeman Carla Mottini Francesca Paolini Michela DAscanio Alberto Ricci Agostino Tafuri Paolo Marchetti Arianna Di Napoli Luciano De Biase Andrea Negro Christian Napoli Paolo Anibaldi Valentina Salvati Darragh Duffy Benjamin Terrier Maurizio Fanciulli Carlo Capalbo Salvatore Sciacchitano Giovanni Blandino Giulia Piaggio Rita Mancini Gennaro Ciliberto 《Cell death & disease》2021,12(11)
964.
Giuseppe Vecchio Francesca Zambianchi Paola Zacchetti Francesco Secundo Giacomo Carrea 《Biotechnology and bioengineering》1999,64(5):545-551
Fourier‐transform infrared (FT‐IR) spectroscopy was employed to investigate potential lyophilization‐induced changes in the secondary structure of lipases from Candida antarctica B and Pseudomonas cepacia. The secondary structure elements were determined by curve fitting of the amide III bands of the two lipases in the lyophilized state in KBr pellets and in solution. It was found that lyophilization decreased the α‐helix and increased the β‐sheet content. However, FT‐IR analysis of crosslinked enzyme crystals of Pseudomonas cepacia lipase also indicated an increase in the β‐sheet content, which appears despite the fact that the enzyme, being in the crystallized state, should possess native conformation. This result partially questions the suitability of FT‐IR for analysis of the structure of solid proteins, at least as far as the β‐sheet content is concerned, because it is possible that the method overestimates the β‐sheets by measuring other hydrogen‐bonded nonperiodic intermolecular structures. No significant modification was observed when lipase from Pseudomonas cepacia was lyophilized in the presence of methoxypoly(ethylene glycol). © 1999 John Wiley & Sons, Inc. Biotechnol Bioeng 64: 545–551, 1999. 相似文献
965.
Adrian Huber Salvatore Demartis Dario Neri 《Journal of molecular recognition : JMR》1999,12(3):198-216
Recently developed scientific instrumentation featuring surface plasmon resonance detection allows the detection of biomolecular interactions in real time and without chemical modification of the binding partners. These biosensors are proving invaluable tools in protein engineering, particularly in research aimed at the isolation and improvement of protein binders and catalysts from macromolecular repertoires containing billions of individual members. This article reviews the use of biosensor technology for the isolation and characterization of engineered antibodies and enzymes. Copyright © 1999 John Wiley & Sons, Ltd. 相似文献
966.
Luca M. Neri Yves Raymond Antonio Giordano S. Capitani Alberto M. Martelli 《Journal of cellular physiology》1999,178(3):284-295
Nuclear lamins are the most abundant components of the nuclear lamina, a 10–50-nm-thick fibrous layer underlying the inner nuclear envelope membrane. Nevertheless, a number of recent investigations performed on epithelial and fibroblast cells have suggested that nuclear lamins are also present within the nucleoplasm and could be important constituents of the nucleoskeleton. We have studied the subnuclear distribution of lamins A and B1 in human erythroleukemia cells by using immunoblotting analysis and immunofluorescent staining of fractionated nuclei. In intact cells and isolated nuclei, antibodies to lamins A and B1 mainly stained the nuclear periphery, although some immunoreactivity was detected in the nuclear interior. However, when chromatin was removed by nuclease digestion and extraction with nonionic detergent or solutions of high ionic strength, a previously masked immunoreactivity for lamin A, but not for lamin B1, became evident in the internal part of the residual structures representing the nuclear matrix or scaffold. Preferential localization of lamin A to the inner part of the nucleus was also demonstrated by the presence of the majority of lamin A in the solubilized inner nuclear network subfraction. In contrast, lamin B1 was mainly recovered in the fraction corresponding to the nuclear periphery. Double labeling experiments showed that lamin A, but not lamin B1, colocalized with coiled and GATA-1 bodies. Thus, our results support the hypothesis that lamin A, but not lamin B1, may be a component of an internal nucleoskeleton in human erythroleukemia cells. J. Cell. Physiol. 178:284–295, 1999. © 1999 Wiley-Liss, Inc. 相似文献
967.
Francesca Resentini Cristina Ruberti Matteo Grenzi Maria Cristina Bonza Alex Costa 《Plant physiology》2021,187(4):1985
Recent insights about the transport mechanisms involved in the in and out of calcium ions in plant organelles, and their role in the regulation of cytosolic calcium homeostasis in different signaling pathways.The transport of Ca2+ across the membranes of subcellular compartments contributes to cytosolic Ca2+ homeostasis as well as environmental and developmental responses. 相似文献
968.
Claudia Giannetto Matteo Gianesella Francesca Arfuso Vincenzo Carcangiu Maria Rizzo Francesco Fazio 《Biological Rhythm Research》2017,48(6):931-938
The aim of the present study was to monitor the daily rhythm of rectal and cutaneous temperatures together with the changes in the serum levels of mitochondrial uncoupling protein 1 (UCP1) in equine and goat species. On five gelding horses and five female goat rectal and cutaneous temperatures were recorded at 4 h intervals for a 48 h period. At the same time points blood samples were collected from each animal. Daily rhythms of rectal and cutaneous temperatures were observed in both species, in both day of monitoring. Mesor value of rectal temperature was statistically higher of about 4 °C than cutaneous temperature, and acrophase was postponed of about 2 h in both day of monitoring and in both species. UCP1 did not show daily rhythmicity in horses and goats. We can speculate that the thermogenesis due to thermal system is auxiliary to keeping the body temperature daily rhythm. 相似文献
969.
Sabrina Berkamp Sang Ho Park Anna A. De Angelis Francesca M. Marassi Stanley J. Opella 《Journal of biomolecular NMR》2017,69(3):111-121
The structure of monomeric human chemokine IL-8 (residues 1–66) was determined in aqueous solution by NMR spectroscopy. The structure of the monomer is similar to that of each subunit in the dimeric full-length protein (residues 1–72), with the main differences being the location of the N-loop (residues 10–22) relative to the C-terminal α-helix and the position of the side chain of phenylalanine 65 near the truncated dimerization interface (residues 67–72). NMR was used to analyze the interactions of monomeric IL-8 (1–66) with ND-CXCR1 (residues 1–38), a soluble polypeptide corresponding to the N-terminal portion of the ligand binding site (Binding Site-I) of the chemokine receptor CXCR1 in aqueous solution, and with 1TM-CXCR1 (residues 1–72), a membrane-associated polypeptide that includes the same N-terminal portion of the binding site, the first trans-membrane helix, and the first intracellular loop of the receptor in nanodiscs. The presence of neither the first transmembrane helix of the receptor nor the lipid bilayer significantly affected the interactions of IL-8 with Binding Site-I of CXCR1. 相似文献
970.
Francesca Verones Stephan Pfister Rosalie van Zelm Stefanie Hellweg 《The International Journal of Life Cycle Assessment》2017,22(8):1247-1256