AUTHOR=Alviz-Gazitua Pablo , Fuentes-Alburquenque Sebastián , Rojas Luis A. , Turner Raymond J. , Guiliani Nicolas , Seeger Michael 

TITLE=The Response of Cupriavidus metallidurans CH34 to Cadmium Involves Inhibition of the Initiation of Biofilm Formation, Decrease in Intracellular c-di-GMP Levels, and a Novel Metal Regulated Phosphodiesterase

JOURNAL=Frontiers in Microbiology

VOLUME=10

YEAR=2019

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2019.01499

DOI=10.3389/fmicb.2019.01499

ISSN=1664-302X

ABSTRACT=<p>Cadmium is a highly toxic heavy metal for biological systems. <italic>Cupriavidus metallidurans</italic> CH34 is a model strain to study heavy metal resistance and bioremediation as it is able to deal with high heavy metal concentrations. Biofilm formation by bacteria is mediated by the second messenger <italic>bis</italic>-(3′–5′)-cyclic dimeric guanosine monophosphate (c-di-GMP). The aim of this study was to characterize the response of <italic>C. metallidurans</italic> CH34 planktonic and biofilm cells to cadmium including their c-di-GMP regulatory pathway. Inhibition of the initiation of biofilm formation and EPS production by <italic>C. metallidurans</italic> CH34 correlates with increased concentration of cadmium. Planktonic and biofilm cells showed similar tolerance to cadmium. During exposure to cadmium an acute decrease of c-di-GMP levels in planktonic and biofilm cells was observed. Transcription analysis by RT-qPCR showed that cadmium exposure to planktonic and biofilm cells induced the expression of the <italic>urf2</italic> gene and the mercuric reductase encoding <italic>merA</italic> gene, which belong to the Tn<italic>501</italic>/Tn<italic>21 mer</italic> operon. After exposure to cadmium, the <italic>cadA</italic> gene involved in cadmium resistance was equally upregulated in both lifestyles. Bioinformatic analysis and complementation assays indicated that the protein encoded by the <italic>urf2</italic> gene is a functional phosphodiesterase (PDE) involved in the c-di-GMP metabolism. We propose to rename the <italic>urf2</italic> gene as <italic>mrp</italic> gene for <underline>m</underline>etal <underline>r</underline>egulated <underline>P</underline>DE. An increase of the second messenger c-di-GMP content by the heterologous expression of the constitutively active diguanylate cyclase PleD correlated with an increase in biofilm formation and cadmium susceptibility. These results indicate that the response to cadmium in <italic>C. metallidurans</italic> CH34 inhibits the initiation of biofilm lifestyle and involves a decrease in c-di-GMP levels and a novel metal regulated PDE.</p>