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Title: Zn 2+-Inducible Expression Platform for Synechococcus sp. Strain PCC 7002 Based on the smtA Promoter/Operator and smtB Repressor

Abstract

Synechococcus sp. strain PCC 7002 has been gaining significance as both a model system for photosynthesis research and for industrial applications. Until recently, the genetic toolbox for this model cyanobacterium was rather limited and relied primarily on tools that only allowed constitutive gene expression. This work describes a two-plasmid, Zn 2+-inducible expression platform that is coupled with a zurA mutation, providing enhanced Zn 2+uptake. The control elements are based on the metal homeostasis system of a class II metallothionein gene ( smtA 7942) and its cognate SmtB 7942repressor fromSynechococcus elongatusstrain PCC 7942. Under optimal induction conditions, yellow fluorescent protein (YFP) levels were about half of those obtained with the strong, constitutive phycocyanin ( cpcBA 6803) promoter ofSynechocystissp. strain PCC 6803. This metal-inducible expression system inSynechococcussp. strain PCC 7002 allowed the titratable gene expression of YFP that was up to 19-fold greater than the background level. This system was utilized successfully to control the expression of theDrosophila melanogasterβ-carotene 15,15′-dioxygenase, NinaB, which is toxic when constitutively expressed from a strong promoter inSynechococcussp. strain PCC 7002. Together, these properties establish this metal-inducible system as an additional useful tool that is capable of controlling gene expression for applications ranging from basic research to syntheticmore » biology inSynechococcussp. strain PCC 7002. IMPORTANCEThis is the first metal-responsive expression system in cyanobacteria, to our knowledge, that does not exhibit low sensitivity for induction, which is one of the major hurdles for utilizing this class of genetic tools. In addition, high levels of expression can be generated that approximate those of established constitutive systems, with the added advantage of titratable control. Together, these properties establish this Zn 2+-inducible system, which is based on the smtA 7942operator/promoter and smtB 7942repressor, as a versatile gene expression platform that expands the genetic toolbox ofSynechococcussp. strain PCC 7002.« less

Authors:
 [1];  [1];  [1];  [1];  [1];  [1]; ORCiD logo [2]
  1. Pennsylvania State Univ., University Park, PA (United States)
  2. Pennsylvania State Univ., University Park, PA (United States); Montana State Univ., Bozeman, MT (United States)
Publication Date:
Research Org.:
Pennsylvania State Univ., University Park, PA (United States)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
1536858
Grant/Contract Number:  
FG02-05ER46222
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Applied and Environmental Microbiology
Additional Journal Information:
Journal Volume: 83; Journal Issue: 3; Journal ID: ISSN 0099-2240
Publisher:
American Society for Microbiology
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; Biotechnology & Applied Microbiology; Microbiology

Citation Formats

Pérez, Adam A., Gajewski, John P., Ferlez, Bryan H., Ludwig, Marcus, Baker, Carol S., Golbeck, John H., and Bryant, Donald A. Zn2+-Inducible Expression Platform for Synechococcus sp. Strain PCC 7002 Based on the smtA Promoter/Operator and smtB Repressor. United States: N. p., 2017. Web. doi:10.1128/aem.02491-16.
Pérez, Adam A., Gajewski, John P., Ferlez, Bryan H., Ludwig, Marcus, Baker, Carol S., Golbeck, John H., & Bryant, Donald A. Zn2+-Inducible Expression Platform for Synechococcus sp. Strain PCC 7002 Based on the smtA Promoter/Operator and smtB Repressor. United States. doi:10.1128/aem.02491-16.
Pérez, Adam A., Gajewski, John P., Ferlez, Bryan H., Ludwig, Marcus, Baker, Carol S., Golbeck, John H., and Bryant, Donald A. Tue . "Zn2+-Inducible Expression Platform for Synechococcus sp. Strain PCC 7002 Based on the smtA Promoter/Operator and smtB Repressor". United States. doi:10.1128/aem.02491-16. https://www.osti.gov/servlets/purl/1536858.
@article{osti_1536858,
title = {Zn2+-Inducible Expression Platform for Synechococcus sp. Strain PCC 7002 Based on the smtA Promoter/Operator and smtB Repressor},
author = {Pérez, Adam A. and Gajewski, John P. and Ferlez, Bryan H. and Ludwig, Marcus and Baker, Carol S. and Golbeck, John H. and Bryant, Donald A.},
abstractNote = {Synechococcus sp. strain PCC 7002 has been gaining significance as both a model system for photosynthesis research and for industrial applications. Until recently, the genetic toolbox for this model cyanobacterium was rather limited and relied primarily on tools that only allowed constitutive gene expression. This work describes a two-plasmid, Zn2+-inducible expression platform that is coupled with a zurA mutation, providing enhanced Zn2+uptake. The control elements are based on the metal homeostasis system of a class II metallothionein gene (smtA7942) and its cognate SmtB7942repressor fromSynechococcus elongatusstrain PCC 7942. Under optimal induction conditions, yellow fluorescent protein (YFP) levels were about half of those obtained with the strong, constitutive phycocyanin (cpcBA6803) promoter ofSynechocystissp. strain PCC 6803. This metal-inducible expression system inSynechococcussp. strain PCC 7002 allowed the titratable gene expression of YFP that was up to 19-fold greater than the background level. This system was utilized successfully to control the expression of theDrosophila melanogasterβ-carotene 15,15′-dioxygenase, NinaB, which is toxic when constitutively expressed from a strong promoter inSynechococcussp. strain PCC 7002. Together, these properties establish this metal-inducible system as an additional useful tool that is capable of controlling gene expression for applications ranging from basic research to synthetic biology inSynechococcussp. strain PCC 7002. IMPORTANCEThis is the first metal-responsive expression system in cyanobacteria, to our knowledge, that does not exhibit low sensitivity for induction, which is one of the major hurdles for utilizing this class of genetic tools. In addition, high levels of expression can be generated that approximate those of established constitutive systems, with the added advantage of titratable control. Together, these properties establish this Zn2+-inducible system, which is based on thesmtA7942operator/promoter and smtB7942repressor, as a versatile gene expression platform that expands the genetic toolbox ofSynechococcussp. strain PCC 7002.},
doi = {10.1128/aem.02491-16},
journal = {Applied and Environmental Microbiology},
issn = {0099-2240},
number = 3,
volume = 83,
place = {United States},
year = {2017},
month = {1}
}

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Works referenced in this record:

The SmtB/ArsR family of metalloregulatory transcriptional repressors: structural insights into prokaryotic metal resistance
journal, June 2003


Unlocking the Constraints of Cyanobacterial Productivity: Acclimations Enabling Ultrafast Growth
journal, July 2016


Bacteriochlorophyll f: properties of chlorosomes containing the “forbidden chlorophyll”
journal, August 2012


Ethylene Synthesis and Regulated Expression of Recombinant Protein in Synechocystis sp. PCC 6803
journal, November 2012


Analysis of the blind Drosophila mutant ninaB identifies the gene encoding the key enzyme for vitamin A formation invivo
journal, January 2001

  • von Lintig, J.
  • Proceedings of the National Academy of Sciences, Vol. 98, Issue 3
  • DOI: 10.1073/pnas.031576398

Use of differential fluorescence induction and optical trapping to isolate environmentally induced genes
journal, June 2001


Zn, Cu and Co in cyanobacteria: selective control of metal availability
journal, June 2003

  • Cavet, Jennifer S.; Borrelly, Gilles P. M.; Robinson, Nigel J.
  • FEMS Microbiology Reviews, Vol. 27, Issue 2-3
  • DOI: 10.1016/S0168-6445(03)00050-0

Synthetic biology of cyanobacteria: unique challenges and opportunities
journal, January 2013


Comparative analysis of idiA and isiA transcription under iron starvation and oxidative stress in Synechococcus elongatus PCC 7942 wild-type and selected mutants
journal, December 2003

  • Yousef, Naeima; Pistorius, Elfriede K.; Michel, Klaus-Peter
  • Archives of Microbiology, Vol. 180, Issue 6
  • DOI: 10.1007/s00203-003-0618-4

Network analysis of transcriptomics expands regulatory landscapes in Synechococcus sp. PCC 7002
journal, August 2016

  • McClure, Ryan S.; Overall, Christopher C.; McDermott, Jason E.
  • Nucleic Acids Research, Vol. 44, Issue 18
  • DOI: 10.1093/nar/gkw737

Cyanobacteria carrying ansmt-lux transcriptional fusion as biosensors for the detection of heavy metal cations
journal, August 1996

  • Erbe, J. L.; Adams, A. C.; Taylor, K. B.
  • Journal of Industrial Microbiology, Vol. 17, Issue 2
  • DOI: 10.1007/BF01570047

Transformation in Agmenellum quadruplicatum
journal, October 1980

  • Stevens, S. E.; Porter, R. D.
  • Proceedings of the National Academy of Sciences, Vol. 77, Issue 10
  • DOI: 10.1073/pnas.77.10.6052

Characterization of psal and psaL Mutants of Synechococcus sp. Strain PCC 7002: A New Model for State Transitions in Cyanobacteria
journal, July 1996


Discovery of a super-strong promoter enables efficient production of heterologous proteins in cyanobacteria
journal, March 2014

  • Zhou, Jie; Zhang, Haifeng; Meng, Hengkai
  • Scientific Reports, Vol. 4, Issue 1
  • DOI: 10.1038/srep04500

Isolation of a prokaryotic metallothionein locus and analysis of transcriptional control by trace metal ions
journal, January 1993


Generic Assignments, Strain Histories and Properties of Pure Cultures of Cyanobacteria
journal, March 1979

  • Stanier, Roger Y.; Deruelles, Josette; Rippka, Rosmarie
  • Microbiology, Vol. 111, Issue 1, p. 1-61
  • DOI: 10.1099/00221287-111-1-1

Group 3 sigma factors in the marine cyanobacterium Synechococcus sp. strain PCC 7002 are required for growth at low temperature
journal, January 2007

  • Inoue-Sakamoto, Kaori; Gruber, Tanja M.; Christensen, Suzanne K.
  • The Journal of General and Applied Microbiology, Vol. 53, Issue 2
  • DOI: 10.2323/jgam.53.89

Structural Determinants of Metal Selectivity in Prokaryotic Metal-responsive Transcriptional Regulators
journal, August 2005


Fur-type transcriptional repressors and metal homeostasis in the cyanobacterium Synechococcus sp. PCC 7002
journal, October 2015


Prokaryotic carbonic anhydrases
journal, October 2000


Vipp1 Is Essential for the Biogenesis of Photosystem I but Not Thylakoid Membranes in Synechococcus sp. PCC 7002
journal, April 2014

  • Zhang, Shuyi; Shen, Gaozhong; Li, Zhongkui
  • Journal of Biological Chemistry, Vol. 289, Issue 23
  • DOI: 10.1074/jbc.M114.555631

SDS-Polyacrylamide Gel Electrophoresis (SDS-PAGE) of Proteins
journal, August 2011


Coordination Chemistry of Bacterial Metal Transport and Sensing
journal, October 2009

  • Ma, Zhen; Jacobsen, Faith E.; Giedroc, David P.
  • Chemical Reviews, Vol. 109, Issue 10
  • DOI: 10.1021/cr900077w

Assembly of Photosystem I: I. INACTIVATION OF THE
journal, March 2002

  • Shen, Gaozhong; Zhao, Jindong; Reimer, Susan K.
  • Journal of Biological Chemistry, Vol. 277, Issue 23
  • DOI: 10.1074/jbc.M201103200

Multiple bacteria encode metallothioneins and SmtA-like zinc fingers: Bacterial metallothioneins and zinc fingers
journal, September 2002


Plastocyanin: Structural and functional analysis
journal, February 1994

  • Redinbo, Matthew R.; Yeates, Todd O.; Merchant, Sabeeha
  • Journal of Bioenergetics and Biomembranes, Vol. 26, Issue 1
  • DOI: 10.1007/BF00763219

Expression of holo-proteorhodopsin in Synechocystis sp. PCC 6803
journal, May 2016


SmtB is a metal-dependent repressor of the cyanobacterial metallothionein gene smtA : identification of a Zn inhibited DNA-protein complex
journal, January 1993

  • Morby, Andrew P.; Turner, Jennifer S.; Huckle, James W.
  • Nucleic Acids Research, Vol. 21, Issue 4
  • DOI: 10.1093/nar/21.4.921

An SmtB-like repressor from Synechocystis PCC 6803 regulates a zinc exporter
journal, September 1998

  • Thelwell, C.; Robinson, N. J.; Turner-Cavet, J. S.
  • Proceedings of the National Academy of Sciences, Vol. 95, Issue 18
  • DOI: 10.1073/pnas.95.18.10728

Chemical rescue of a site-modified ligand to a [4Fe–4S] cluster in PsaC, a bacterial-like dicluster ferredoxin bound to Photosystem I
journal, June 2007

  • Antonkine, Mikhail L.; Maes, Estelle M.; Czernuszewicz, Roman S.
  • Biochimica et Biophysica Acta (BBA) - Bioenergetics, Vol. 1767, Issue 6
  • DOI: 10.1016/j.bbabio.2007.02.003

Evidence for genetic transformation in blue-green alga Anacystis nidulans
journal, January 1970

  • Shestakov, S. V.; Khyen, Nguyen Than
  • MGG Molecular & General Genetics, Vol. 107, Issue 4
  • DOI: 10.1007/BF00441199

Synthetic Biology Toolbox for Controlling Gene Expression in the Cyanobacterium Synechococcus sp. strain PCC 7002
journal, September 2014

  • Markley, Andrew L.; Begemann, Matthew B.; Clarke, Ryan E.
  • ACS Synthetic Biology, Vol. 4, Issue 5
  • DOI: 10.1021/sb500260k

Development of a Univariate Membrane-Based Mid-Infrared Method for Protein Quantitation and Total Lipid Content Analysis of Biological Samples
journal, January 2014

  • Strug, Ivona; Utzat, Christopher; Cappione, Amedeo
  • Journal of Analytical Methods in Chemistry, Vol. 2014
  • DOI: 10.1155/2014/657079

A Drosophila Complementary DNA Resource
journal, March 2000


Luminescent Whole-Cell Cyanobacterial Bioreporter for Measuring Fe Availability in Diverse Marine Environments
journal, December 2006

  • Boyanapalli, R.; Bullerjahn, G. S.; Pohl, C.
  • Applied and Environmental Microbiology, Vol. 73, Issue 3
  • DOI: 10.1128/AEM.01670-06

The Production of Hydrogen Peroxide by Blue-Green Algae: a Survey
journal, December 1973