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R Díaz Orejas

Publications and source records attributed to R Díaz Orejas.

2 recordsLinked to original sources
PubMed1998

Functional interactions between chpB and parD, two homologous conditional killer systems found in the Escherichia coli chromosome and in plasmid R1.

parD and chpB are homologous conditional killer systems of plasmid and chromosomal origin, respectively. They are bicistronic operons encoding a killer protein (Kid and ChpBK) and an antidote (Kis and ChpBI). It is shown that the antidote of the chpB system can neutralize the toxin of the parD system. This activity is improved by particular amino acid changes at the amino end of the ChpBI antidote. It is further shown that the chpB system is weakly autoregulated and that the activity of a second promoter, previously identified upstream of the regulated promoter, can modulate the functional interactions between the chpB and parD systems.

Amino Acid Sequence↗
PubMed1994

Phage P4 DNA replication in vitro.

Phage P4 DNA is replicated in cell-free extracts of Escherichia coli in the presence of partially purified P4 alpha protein [Krevolin and Calendar (1985), J. Mol. Biol. 182, 507-517]. Using a modified in vitro replication assay, we have further characterized this process. Analysis by agarose gel electrophoresis and autoradiography of in vitro replicated molecules demonstrates that the system yields supercoiled monomeric DNA as the main product. Electron microscopic analysis of in vitro generated intermediates indicates that DNA synthesis initiates in vitro mainly at ori, the origin of replication used in vivo. Replication proceeds from this origin bidirectionally, resulting in theta-type molecules. In contrast to the in vivo situation, no extensive single-stranded regions were found in these intermediates. The initiation proteins of the host, DnaB and DnaG, and the chaperones DnaJ and DnaK are not required for P4 replication, because polyclonal antibodies against those polypeptides do not inhibit the process. The reaction is inhibited by antibodies against the SSB protein, and by ara-CTP, a specific inhibitor of DNA polymerase III holoenzyme. Consistent with previous reports, P4 in vitro replication is independent of transcription by host RNA polymerase. Novobiocin, a DNA gyrase inhibitor, strongly inhibits P4 DNA synthesis, indicating that form I DNA is the required substrate.

Base Sequence↗