TY - JOUR
T1 - Copper-responsive gene expression in the methanotroph
T2 - Methylosinus trichosporium OB3b
AU - Kenney, Grace E.
AU - Sadek, Monica
AU - Rosenzweig, Amy C.
N1 - Funding Information:
This work was supported by NSF grant MCB0842366 and the Institute for Sustainability and Energy at Northwestern (ISEN). G. E. K. was supported by American Heart Association predoctoral fellowship 14PRE20460104 and the Northwestern University Presidential Fellowship. Imaging work was performed at the Biological Imaging Facility generously supported by the Office for Research (Northwestern University). The qRT-PCR experiments were performed at the High Throughput Analysis Facility, supported by the Robert H. Lurie Cancer Center and the Office for Research (Northwestern University). Nucleic acid quality control and genome sequencing were performed with the help of the NUSeq Core Facility (Northwestern University.) Mass spectrometry was performed at the Integrated Molecular Structure Education and Research Center. We additionally thank Dr. George Georgiou for permission to use the PP358 strain (sourced from ATCC) and Dr. Stefan Green (University of Illinois, Chicago) for assistance with the PP358 genome.
Publisher Copyright:
© 2016 The Royal Society of Chemistry.
PY - 2016/9
Y1 - 2016/9
N2 - Methanotrophic bacteria convert methane to methanol using methane monooxygenase (MMO) enzymes. In many strains, either an iron-containing soluble (sMMO) or a copper-containing particulate (pMMO) enzyme can be produced depending on copper availability; the mechanism of this copper switch has not been elucidated. A key player in methanotroph copper homeostasis is methanobactin (Mbn), a ribosomally produced, post-translationally modified natural product with a high affinity for copper. The Mbn precursor peptide is encoded within an operon that contains a range of putative transporters, regulators, and biosynthetic proteins, but the involvement of these genes in Mbn-related processes remains unclear. Extensive time-dependent qRT-PCR studies of Methylosinus trichosporium OB3b and the constitutive sMMO-producing mutant M. trichosporium OB3b PP358 show that the Mbn operon is indeed copper-regulated, providing experimental support for its bioinformatics-based identification. Moreover, the Mbn operon is co-regulated with the sMMO operon and reciprocally regulated with the pMMO operon. Within the Mbn and sMMO operons, a subset of regulatory genes exhibits a distinct and shared pattern of expression, consistent with their proposed functions as internal regulators. In addition, genome sequencing of the M. trichosporium OB3b PP358 mutant provides new evidence for the involvement of genes adjacent to the pMMO operon in methanotroph copper homeostasis.
AB - Methanotrophic bacteria convert methane to methanol using methane monooxygenase (MMO) enzymes. In many strains, either an iron-containing soluble (sMMO) or a copper-containing particulate (pMMO) enzyme can be produced depending on copper availability; the mechanism of this copper switch has not been elucidated. A key player in methanotroph copper homeostasis is methanobactin (Mbn), a ribosomally produced, post-translationally modified natural product with a high affinity for copper. The Mbn precursor peptide is encoded within an operon that contains a range of putative transporters, regulators, and biosynthetic proteins, but the involvement of these genes in Mbn-related processes remains unclear. Extensive time-dependent qRT-PCR studies of Methylosinus trichosporium OB3b and the constitutive sMMO-producing mutant M. trichosporium OB3b PP358 show that the Mbn operon is indeed copper-regulated, providing experimental support for its bioinformatics-based identification. Moreover, the Mbn operon is co-regulated with the sMMO operon and reciprocally regulated with the pMMO operon. Within the Mbn and sMMO operons, a subset of regulatory genes exhibits a distinct and shared pattern of expression, consistent with their proposed functions as internal regulators. In addition, genome sequencing of the M. trichosporium OB3b PP358 mutant provides new evidence for the involvement of genes adjacent to the pMMO operon in methanotroph copper homeostasis.
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U2 - 10.1039/c5mt00289c
DO - 10.1039/c5mt00289c
M3 - Article
C2 - 27087171
AN - SCOPUS:84988419934
SN - 1756-5901
VL - 8
SP - 931
EP - 940
JO - Metallomics
JF - Metallomics
IS - 9
ER -