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McCallum, M. E.

Publications and source records attributed to McCallum, M. E..

2 recordsLinked to original sources

Quorum sensing mediates morphology and motility transitions in the model archaeon Haloferax volcanii

Quorum sensing (QS) is a mechanism of intercellular communication that enables microbes to alter gene expression and adapt to the environment. This cell-cell signaling is necessary for intra- and interspecies behaviors such as virulence and biofilm formation. While QS has been extensively studied in bacteria, little is known about cell-cell communication in archaea. Here we established an archaeal model system to study QS. We showed that for Haloferax volcanii, the transition from motile rods to non-motile disks is dependent on a possibly novel, secreted small molecule present in cell-free conditioned medium (CM). Moreover, we determined that this putative QS molecule fails to induce the morphology transition in mutants lacking the regulatory factors, DdfA and CirA. Using quantitative proteomics of wild-type cells, we detected significant differential abundances of 236 proteins in the presence of CM. Conversely, in the {Delta}ddfA mutant, addition of CM resulted in only 110 proteins of significant differential abundances. These results confirm that DdfA is involved in CM-dependent regulation. CirA, along with other proteins involved in morphology and swimming motility transitions, is among the proteins regulated by DdfA. These discoveries significantly advance our understanding of microbial communication within archaeal species.

microbiology↗

The L-alanosine gene cluster encodes a pathway for diazeniumdiolate biosynthesis

N-nitroso-containing natural products are bioactive metabolites with antibacterial and anticancer properties. In particular, compounds containing the diazeniumdiolate (N-nitrosohydroxylamine) group display a wide range of bioactivities ranging from cytotoxicity to metal chelation. Despite the importance of this structural motif, knowledge of its biosynthesis is limited. Herein, we describe the discovery of a biosynthetic gene cluster in Streptomyces alanosinicus ATCC 15710 responsible for producing the diazeniumdiolate natural product O_SCPCAPLC_SCPCAP-alanosine. Gene disruption and stable isotope feeding experiments identified essential biosynthetic genes and revealed the nitrogen source of the N-nitroso group. Additional biochemical characterization of the biosynthetic enzymes revealed that the non-proteinogenic amino acid O_SCPCAPLC_SCPCAP-2,3-diaminopropionic acid (O_SCPCAPLC_SCPCAP-Dap) is synthesized and loaded onto a peptidyl carrier protein (PCP) domain in O_SCPCAPLC_SCPCAP-alanosine biosynthesis, which we propose may be a mechanism of handling unstable intermediates generated en route to the diazeniumdiolate. This research framework will facilitate efforts to determine the biochemistry of diazeniumdiolate formation.

biochemistry↗