Developing a methodology for the National Frog Survey of Ireland: a pilot study in Co. Mayo.

A National Frog Survey of Ireland is planned for spring 2011. We conducted a pilot survey of 25 water bodies in ten 0.25 km2 survey squares in Co. Mayo during spring 2010. Drainage ditches were the most commonly available site for breeding and, generally, two 100 m stretches of ditch were surveyed in each square. The restricted period for peak spawning activity renders any methodology utilizing only one site visit inherently risky. Consequently, each site was visited three times from late March to early April. Occurrence of spawn declined significantly from 72 % to 44 % between the first and third visit whilst the overall occurrence of spawn at all sites was 76 %. As the breeding season advanced, spawn either hatched or was predated and, therefore, disappeared. In those water bodies where spawning was late, however, greater densities of spawn were deposited than in those sites where breeding was early. Consequently, spawn density and estimated frog density did not differ significantly between site visits. Future surveys should nevertheless include multiple site visits to avoid biased estimation of species occurrence and distribution. Ecological succession was identified as the main threat present at 44 % of sites.

MarA-mediated transcriptional repression of the rob promoter

The Escherichia coli transcriptional regulator MarA affects functions that include antibiotic resistance, persistence, and survival. MarA functions as an activator or repressor of transcription utilizing similar degenerate DNA sequences (marboxes) with three different binding site configurations with respect to the RNA polymerase-binding sites. We demonstrate that MarA down-regulates rob transcripts both in vivo and in vitro via a MarA-binding site within the rob promoter that is positioned between the -10 and -35 hexamers. As for the hdeA and purA promoters, which are repressed by MarA, the rob marbox is also in the "backward" orientation. Protein-DNA interactions show that SoxS and Rob, like MarA, bind the same marbox in the rob promoter. Electrophoretic mobility shift analyses with a MarA-specific antibody demonstrate that MarA and RNA polymerase form a ternary complex with the rob promoter DNA. Transcription experiments in vitro and potassium permanganate footprinting analysis show that MarA affects the RNA polymerase-mediated closed to open complex formation at the rob promoter.

The Escherichia coli transcriptional regulator MarA directly represses transcription of purA and hdeA

The Escherichia coli MarA protein mediates a response to multiple environmental stresses through the activation or repression in vivo of a large number of chromosomal genes. Transcriptional activation for a number of these genes has been shown to occur via direct interaction of MarA with a 20-bp degenerate asymmetric "marbox" sequence. It was not known whether repression by MarA was also direct. We found that purified MarA was sufficient in vitro to repress transcription of both purA and hdeA. Transcription and electrophoretic mobility shift experiments in vitro using mutant promoters suggested that the marbox involved in the repression overlapped the -35 promoter motif and was in the "backward" orientation. This organization contrasts with that of the class II promoters activated by MarA, in which the marbox also overlaps the -35 motif but is in the "forward" orientation. We conclude that MarA, a member of the AraC/XylS family, can act directly as a repressor or an activator, depending on the position and orientation of the marbox within a promoter.