Double-stranded DMA Replication Intermediates in Human Malignant Melanoma Cells1
نویسندگان
چکیده
DNA synthesis was investigated in human malignant mela noma cells. The DNA was extracted in a neutral nondenaturing buffer at 25°and characterized by agarose gel electrophoresis. Radioactive DNA pulse labeled for 10 to 30 min was released from the chromosomes in the form of double-stranded frag ments. The release of the fragments from the chromosomes is prevented by lysing the cells at 0°instead of at 25°suggesting that the release is enzymatic. The size of the fragments ranges between 4.6 and 5.6 x 106 daltons. After a time lag, the fragments are joined together to produce a high-molecularweight DNA.
منابع مشابه
Break-induced replication is a source of mutation clusters underlying kataegis.
Clusters of simultaneous multiple mutations can be a source of rapid change during carcinogenesis and evolution. Such mutation clusters have been recently shown to originate from DNA damage within long single-stranded DNA (ssDNA) formed at resected double-strand breaks and dysfunctional replication forks. Here, we identify double-strand break (DSB)-induced replication (BIR) as another powerful ...
متن کاملBlocking single-stranded transferred DNA conversion to double-stranded intermediates by overexpression of yeast DNA REPLICATION FACTOR A.
Agrobacterium tumefaciens delivers its single-stranded transferred DNA (T-strand) into the host cell nucleus, where it can be converted into double-stranded molecules. Various studies have revealed that double-stranded transfer DNA (T-DNA) intermediates can serve as substrates by as yet uncharacterized integration machinery. Nevertheless, the possibility that T-strands are themselves substrates...
متن کاملAction of 3-methylquercetin on poliovirus RNA replication.
3-Methylquercetin is a natural flavone that powerfully blocks poliovirus replication. This compound inhibits selectively poliovirus RNA synthesis both in infected cells and in cell-free systems. Poliovirus double-stranded RNA (replicative forms) is still made in the presence of this inhibitor, whereas the synthesis of single-stranded RNA and the formation of replicative intermediates are drasti...
متن کاملReplicative intermediates of porcine circovirus in animal tissue cultured cells or in bacteria undergoing copy-release replication.
Porcine circovirus (PCV) has been assumed to replicate its genome via the rolling-circle replication (RCR) mechanism because it encodes a Rep protein that contains several amino acid motifs commonly found in other RCR biological systems. Two proteins, Rep and Rep', are essential for PCV DNA replication in mammalian cells. In this work, replicative intermediates of PCV-infected porcine kidney (P...
متن کاملTwo independent molecular pathways for recombinant adeno-associated virus genome conversion occur after UV-C and E4orf6 augmentation of transduction.
Numerous environmental influences have been demonstrated to enhance recombinant adeno-associated virus (rAAV) transduction. Such findings are the foundation of developing new and innovative strategies to improve the efficiency of rAAV as a gene therapy vector. Several of these environmental factors included genotoxic stresses such as UV and y irradiation as well as certain adenoviral gene produ...
متن کامل