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Title: Zam Is a Redox-Regulated Member of the RNB-Family Required for Optimal Photosynthesis in Cyanobacteria

Journal Article · · Microorganisms

The zam gene mediating resistance to acetazolamide in cyanobacteria was discovered thirty years ago during a drug tolerance screen. We use phylogenetics to show that Zam proteins are distributed across cyanobacteria and that they form their own unique clade of the ribonuclease II/R (RNB) family. Despite being RNB family members, multiple sequence alignments reveal that Zam proteins lack conservation and exhibit extreme degeneracy in the canonical active site—raising questions about their cellular function(s). Several known phenotypes arise from the deletion of zam, including drug resistance, slower growth, and altered pigmentation. Using room-temperature and low-temperature fluorescence and absorption spectroscopy, we show that deletion of zam results in decreased phycocyanin synthesis rates, altered PSI:PSII ratios, and an increase in coupling between the phycobilisome and PSII. Conserved cysteines within Zam are identified and assayed for function using in vitro and in vivo methods. We show that these cysteines are essential for Zam function, with mutation of either residue to serine causing phenotypes identical to the deletion of Zam. Redox regulation of Zam activity based on the reversible oxidation-reduction of a disulfide bond involving these cysteine residues could provide a mechanism to integrate the ‘central dogma’ with photosynthesis in cyanobacteria.

Sponsoring Organization:
USDOE
Grant/Contract Number:
SC0018368; SC0020361; SC0019306
OSTI ID:
1869006
Journal Information:
Microorganisms, Journal Name: Microorganisms Journal Issue: 5 Vol. 10; ISSN 2076-2607; ISSN MICRKN
Publisher:
MDPI AGCopyright Statement
Country of Publication:
Switzerland
Language:
English

References (47)

UCSF Chimera?A visualization system for exploratory research and analysis journal January 2004
A protein is involved in accessibility of the inhibitor acetazolamide to the carbonic anhydrase(s) in the cyanobacterium Synechocystis PCC 6803 journal February 1995
Nitrogen chlorosis in blue-green algae journal January 1969
The effects of nitrogen limitation on the ultrastructure of the cyanobacterium Agmenellum quadruplicatum journal November 1981
Genetic and genomic analysis of RNases in model cyanobacteria journal January 2015
A simple method for stabilizing protein-sulfhydryl groups during SDS-gel electrophoresis journal June 1978
Basic local alignment search tool journal October 1990
Chromatophore Genome Sequence of Paulinella Sheds Light on Acquisition of Photosynthesis by Eukaryotes journal March 2008
Characterization of the Functional Domains of Escherichia coli RNase II journal July 2006
An Important Role for RNase R in mRNA Decay journal January 2005
Structure of the Active Subunit of the Yeast Exosome Core, Rrp44: Diverse Modes of Substrate Recruitment in the RNase II Nuclease Family journal March 2008
Unravelling the dynamics of RNA degradation by ribonuclease II and its RNA-bound complex journal September 2006
Enzymatic assembly of DNA molecules up to several hundred kilobases journal April 2009
I-TASSER: a unified platform for automated protein structure and function prediction journal March 2010
Mechanical regulation of photosynthesis in cyanobacteria journal March 2020
Highly accurate protein structure prediction with AlphaFold journal July 2021
The Escherichia coli major exoribonuclease RNase II is a component of the RNA degradosome journal December 2014
Quality control of ribosomal RNA mediated by polynucleotide phosphorylase and RNase R journal May 2003
Complete genome of a nonphotosynthetic cyanobacterium in a diatom reveals recent adaptations to an intracellular lifestyle journal July 2014
Polynucleotide phosphorylase and ribonuclease II are required for cell viability and mRNA turnover in Escherichia coli K-12. journal January 1986
Ssd1p of Saccharomyces cerevisiae Associates with RNA journal June 1997
The vacB Gene Required for Virulence inShigella flexneri and Escherichia coli Encodes the Exoribonuclease RNase R journal June 1998
Exoribonuclease R Interacts with Endoribonuclease E and an RNA Helicase in the Psychrotrophic Bacterium Pseudomonas syringae Lz4W journal April 2005
Proteome-wide Light/Dark Modulation of Thiol Oxidation in Cyanobacteria Revealed by Quantitative Site-specific Redox Proteomics journal August 2014
Inactivation of the RNA helicase CrhR impacts a specific subset of the transcriptome in the cyanobacterium Synechocystis sp. PCC 6803 journal October 2018
The temperature-regulated DEAD-box RNA helicase CrhR interactome: autoregulation and photosynthesis-related transcripts journal September 2021
Comparative sequence analysis of ribonucleases HII, III, II PH and D journal August 1997
Exoribonuclease superfamilies: structural analysis and phylogenetic distribution journal March 2001
RNase II binds to RNase E and modulates its endoribonucleolytic activity in the cyanobacterium Anabaena PCC 7120 journal February 2020
AlphaFold Protein Structure Database: massively expanding the structural coverage of protein-sequence space with high-accuracy models journal November 2021
RNA polyadenylation and degradation in different Archaea; roles of the exosome and RNase R journal October 2006
Structural insights into RNA unwinding and degradation by RNase R journal October 2017
Inactivation of a Low Temperature-Induced RNA Helicase in Synechocystis sp. PCC 6803: Physiological and Morphological Consequences journal February 2012
WebLogo: A Sequence Logo Generator journal May 2004
Redox-Regulated RNA Helicase Expression journal October 2000
Properties of a Mutant from Synechocystis PCC6803 Resistant to Acetazolamide, an Inhibitor of Carbonic Anhydrase journal August 1990
Progress and challenges in engineering cyanobacteria as chassis for light‐driven biotechnology journal August 2019
A protein complex mediating mRNA degradation in Escherichia coli journal November 1994
The Production of Hydrogen Peroxide by Blue-Green Algae: a Survey journal December 1973
The critical role of RNA processing and degradation in the control of gene expression journal September 2010
A single mutation in Escherichia coli ribonuclease II inactivates the enzyme without affecting RNA binding: RNase II mutant with RNA binding but no activity journal December 2004
Life cycle of a cyanobacterial carboxysome journal May 2020
Unicellular Cyanobacterial Distributions Broaden the Oceanic N2 Fixation Domain journal February 2010
REDOX REGULATION: A Broadening Horizon journal June 2005
Nucleic Acid Recognition by OB-Fold Proteins journal June 2003
The rnb Gene of Synechocystis PCC6803 Encodes a RNA Hydrolase Displaying RNase II and Not RNase R Enzymatic Properties journal March 2012
Characterization of protein redox dynamics induced during light-to-dark transitions and nutrient limitation in cyanobacteria journal July 2014