We examined the effects of acute intraocular pressure elevation on the intraretinal electroretinogram in the cat dark-adapted eye. Perfusion pressure of the eye (mean arterial pressure minus intraocular pressure) was regulated within the range of 25-75 mm Hg by increasing the intraocular pressure, which was adjusted by changing the height of the reservoir of balanced salt solution perfused into the eye through a needle placed in the anterior chamber. The vitreal electroretinogram, transepithelial c-wave, slow PIII and light-evoked decrease in extracellular K+ concentration were recorded during intraocular pressure elevation in the dark-adapted retina. The c-wave increased in amplitude between 40 and 75 mm Hg of perfusion pressure and decreased below 40 mm Hg in some cases. These changes were unrelated to the standing potential. The transepithelial c-wave, slow PIII and light-evoked decrease in extracellular K+ concentration were larger in the range of perfusion pressure examined. These suggest that some factor induces electroretinographic changes that are not readily explained by hypoxia or pH changes alone.

Animals↗

PubMedDate not supplied

Escherichia coli and Bacillus subtilis PriA proteins essential for recombination-dependent DNA replication: involvement of ATPase/helicase activity of PriA for inducible stable DNA replication.

The E. coli PriA protein, a DEXH-type DNA helicase with unique zinc finger-like motifs interrupting the helicase domains, is an essential component of the phiX174-type primosome and plays critical roles in RecA-dependent inducible and constitutive stable DNA replication (iSDR and cSDR, respectively) as well as in recombination-dependent repair of double-stranded DNA breaks. B. subtilis PriA (BsPriA) protein contains the conserved helicase domains as well as zinc finger-like motifs with 34% overall identity with the E. coli counterpart. We overexpressed and purified BsPriA and examined its biochemical properties. BsPriA binds specifically to both n'-pas (primosome assembly site) and D-loop and hydrolyzes ATP in the presence of n'-pas albeit with a specific activity about 30% of that of E. coli PriA. However, it is not capable of supporting n'-pas-dependent replication in vitro, nor is it able to support ColE1-type plasmid replication in vivo which requires the function of the phiX174-type primosome. We also show that a zinc finger mutant is not able to support recombination-dependent DNA replication, as measured by the level of iSDR after a period of thymine starvation, nor wild-type level of growth, cell morphology and UV resistance. Unexpectedly, we discovered that an ATPase-deficient mutant (K230D) is not able to support iSDR to a full extent, although it can restore normal growth rate and UV resistance as well as non-filamentous morphology in priA1::kan mutant. K230D was previously reported to be fully functional in assembly of the phiX174-type primosome at a single-stranded n'-pas. Our results indicate that ATP hydrolysis/ helicase activity of PriA may be specifically required for DNA replication from recombination intermediates in vivo.

Adenosine Triphosphatases↗