skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Differential Regulation of the Two Ferrochelatase Paralogues in Shewanella loihica PV-4 in Response to Environmental Stresses

Abstract

Determining the function and regulation of paralogues is important in understanding microbial functional genomics and environmental adaptation. Heme homeostasis is crucial for the survival of environmental microorganisms. MostShewanellaspecies encode two paralogues of ferrochelatase, the terminal enzyme in the heme biosynthesis pathway. The function and transcriptional regulation of two ferrochelatase genes, hemH1 and hemH2, were investigated inShewanellaloihicaPV-4. The disruption of hemH1 but not hemH2 resulted in a significant accumulation of extracellular protoporphyrin IX (PPIX), the precursor to heme, and decreased intracellular heme levels.hemH1 was constitutively expressed, and the expression of hemH2 increased when hemH1 was disrupted. The transcription ofhemH1was regulated by the housekeeping sigma factor RpoD and potentially regulated by OxyR, while hemH2 appeared to be regulated by the oxidative stress-associated sigma factor RpoE2. When an oxidative stress condition was mimicked by adding H 2O 2 to the medium or exposing the culture to light, PPIX accumulation was suppressed in the Δ hemH1 mutant. Consistently, transcriptome analysis indicated enhanced iron uptake and suppressed heme synthesis in the ΔhemH1 mutant. These data indicate that the two paralogues are functional in the heme synthesis pathway but regulated by environmental conditions, providing insights into the understanding of bacterial response to environmental stresses and amore » great potential to commercially produce porphyrin compounds.Shewanella is capable of utilizing a variety of electron acceptors for anaerobic respiration because of the existence of multiple c -type cytochromes in which heme is an essential component. The cytochrome-mediated electron transfer across cellular membranes could potentially be used for biotechnological purposes, such as electricity generation in microbial fuel cells and dye decolorization. However, the mechanism underlying the regulation of biosynthesis of heme and cytochromes is poorly understood. In conclusion, our study has demonstrated that two ferrochelatase genes involved in heme biosynthesis are differentially regulated in response to environmental stresses, including light and reactive oxygen species. This is an excellent example showing how bacteria have evolved to maintain cellular heme homeostasis. More interestingly, the high yields of extracellular protoporphyrin IX by theShewanella loihicaPV-4 mutants could be utilized for commercial production of this valuable chemical via bacterial fermentation.« less

Authors:
 [1];  [2];  [3];  [3];  [3];  [4];  [2];  [2];  [2];  [5];  [6]
  1. Chinese Academy of Sciences (CAS), Wuhan (China). Inst. of Hydrobiology; Univ. of Chinese Academy of Sciences, Beijing (China); Univ. of Oklahoma, Norman, OK (United States). Inst. for Environmental Genomics, Dept. of Microbiology and Plant Biology
  2. Univ. of Oklahoma, Norman, OK (United States). Inst. for Environmental Genomics, Dept. of Microbiology and Plant Biology
  3. Chinese Academy of Sciences (CAS), Wuhan (China). Inst. of Hydrobiology; Univ. of Chinese Academy of Sciences, Beijing (China)
  4. Hubei Engineering Univ., Xiaogan (China). School of Life Sciences and Technology
  5. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Earth Science Division
  6. Univ. of Oklahoma, Norman, OK (United States). Inst. for Environmental Genomics, Dept. of Microbiology and Plant Biology; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Earth Science Division; Tsinghua Univ., Beijing (China). State Key Joint Lab. of Environment Simulation and Pollution Control, School of Environment
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
Sponsoring Org.:
USDOE Office of Science (SC); Chinese Academy of Sciences (CAS)
OSTI Identifier:
1378739
Alternate Identifier(s):
OSTI ID: 1394586
Grant/Contract Number:  
AC02-05CH11231; FG02-07ER64383; Y15103-1-401; AC05-00OR22725
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Applied and Environmental Microbiology
Additional Journal Information:
Journal Volume: 82; Journal Issue: 17; Journal ID: ISSN 0099-2240
Publisher:
American Society for Microbiology
Country of Publication:
United States
Language:
English
Subject:
54 ENVIRONMENTAL SCIENCES; 60 APPLIED LIFE SCIENCES

Citation Formats

Qiu, Dongru, Xie, Ming, Dai, Jingcheng, An, Weixing, Wei, Hehong, Tian, Chunyuan, Kempher, Megan L., Zhou, Aifen, He, Zhili, Gu, Baohua, and Zhou, Jizhong. Differential Regulation of the Two Ferrochelatase Paralogues in Shewanella loihica PV-4 in Response to Environmental Stresses. United States: N. p., 2016. Web. doi:10.1128/AEM.00203-16.
Qiu, Dongru, Xie, Ming, Dai, Jingcheng, An, Weixing, Wei, Hehong, Tian, Chunyuan, Kempher, Megan L., Zhou, Aifen, He, Zhili, Gu, Baohua, & Zhou, Jizhong. Differential Regulation of the Two Ferrochelatase Paralogues in Shewanella loihica PV-4 in Response to Environmental Stresses. United States. doi:10.1128/AEM.00203-16.
Qiu, Dongru, Xie, Ming, Dai, Jingcheng, An, Weixing, Wei, Hehong, Tian, Chunyuan, Kempher, Megan L., Zhou, Aifen, He, Zhili, Gu, Baohua, and Zhou, Jizhong. Fri . "Differential Regulation of the Two Ferrochelatase Paralogues in Shewanella loihica PV-4 in Response to Environmental Stresses". United States. doi:10.1128/AEM.00203-16. https://www.osti.gov/servlets/purl/1378739.
@article{osti_1378739,
title = {Differential Regulation of the Two Ferrochelatase Paralogues in Shewanella loihica PV-4 in Response to Environmental Stresses},
author = {Qiu, Dongru and Xie, Ming and Dai, Jingcheng and An, Weixing and Wei, Hehong and Tian, Chunyuan and Kempher, Megan L. and Zhou, Aifen and He, Zhili and Gu, Baohua and Zhou, Jizhong},
abstractNote = {Determining the function and regulation of paralogues is important in understanding microbial functional genomics and environmental adaptation. Heme homeostasis is crucial for the survival of environmental microorganisms. MostShewanellaspecies encode two paralogues of ferrochelatase, the terminal enzyme in the heme biosynthesis pathway. The function and transcriptional regulation of two ferrochelatase genes, hemH1 and hemH2, were investigated inShewanellaloihicaPV-4. The disruption of hemH1 but not hemH2 resulted in a significant accumulation of extracellular protoporphyrin IX (PPIX), the precursor to heme, and decreased intracellular heme levels.hemH1 was constitutively expressed, and the expression of hemH2 increased when hemH1 was disrupted. The transcription ofhemH1was regulated by the housekeeping sigma factor RpoD and potentially regulated by OxyR, while hemH2 appeared to be regulated by the oxidative stress-associated sigma factor RpoE2. When an oxidative stress condition was mimicked by adding H2O2 to the medium or exposing the culture to light, PPIX accumulation was suppressed in the Δ hemH1 mutant. Consistently, transcriptome analysis indicated enhanced iron uptake and suppressed heme synthesis in the ΔhemH1 mutant. These data indicate that the two paralogues are functional in the heme synthesis pathway but regulated by environmental conditions, providing insights into the understanding of bacterial response to environmental stresses and a great potential to commercially produce porphyrin compounds.Shewanella is capable of utilizing a variety of electron acceptors for anaerobic respiration because of the existence of multiple c -type cytochromes in which heme is an essential component. The cytochrome-mediated electron transfer across cellular membranes could potentially be used for biotechnological purposes, such as electricity generation in microbial fuel cells and dye decolorization. However, the mechanism underlying the regulation of biosynthesis of heme and cytochromes is poorly understood. In conclusion, our study has demonstrated that two ferrochelatase genes involved in heme biosynthesis are differentially regulated in response to environmental stresses, including light and reactive oxygen species. This is an excellent example showing how bacteria have evolved to maintain cellular heme homeostasis. More interestingly, the high yields of extracellular protoporphyrin IX by theShewanella loihicaPV-4 mutants could be utilized for commercial production of this valuable chemical via bacterial fermentation.},
doi = {10.1128/AEM.00203-16},
journal = {Applied and Environmental Microbiology},
issn = {0099-2240},
number = 17,
volume = 82,
place = {United States},
year = {2016},
month = {6}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record

Save / Share:

Works referenced in this record:

Mosaic Origin of the Heme Biosynthesis Pathway in Photosynthetic Eukaryotes
journal, August 2005

  • Oborník, Miroslav; Green, Beverley R.
  • Molecular Biology and Evolution, Vol. 22, Issue 12
  • DOI: 10.1093/molbev/msi230

Genetics, Physiology, and Evolutionary Relationships of the Genus Buchnera: Intracellular Symbionts of Aphids
journal, October 1995


Molecular basis of inherited microcytic anemia due to defects in iron acquisition or heme synthesis
journal, March 2009


The Biochemistry of Heme Synthesis in Porphyria and in the Porphyrinurias
journal, March 1998


A whole genome view of prokaryotic haem biosynthesis
journal, August 2002


Molecular hijacking of siroheme for the synthesis of heme and d1 heme
journal, October 2011

  • Bali, S.; Lawrence, A. D.; Lobo, S. A.
  • Proceedings of the National Academy of Sciences, Vol. 108, Issue 45
  • DOI: 10.1073/pnas.1108228108

Noncanonical coproporphyrin-dependent bacterial heme biosynthesis pathway that does not use protoporphyrin
journal, February 2015

  • Dailey, Harry A.; Gerdes, Svetlana; Dailey, Tamara A.
  • Proceedings of the National Academy of Sciences, Vol. 112, Issue 7
  • DOI: 10.1073/pnas.1416285112

Ecology and Biotechnology of the Genus Shewanella
journal, October 2007


Towards environmental systems biology of Shewanella
journal, July 2008

  • Fredrickson, James K.; Romine, Margaret F.; Beliaev, Alexander S.
  • Nature Reviews Microbiology, Vol. 6, Issue 8
  • DOI: 10.1038/nrmicro1947

Photosensitivity of a protoporphyrin-accumulating, light-sensitive mutant (visA) of Escherichia coli K-12.
journal, December 1991

  • Nakahigashi, K.; Nishimura, K.; Miyamoto, K.
  • Proceedings of the National Academy of Sciences, Vol. 88, Issue 23
  • DOI: 10.1073/pnas.88.23.10520

Accumulation of protoporphyrin IX in light-sensitive mutants of Escherichia coli
journal, October 1992


Photooxidation and Singlet Oxygen Sensitization by Protoporphyrin ix and its Photooxidation Products
journal, October 1982


Overcoming the Heme Paradox: Heme Toxicity and Tolerance in Bacterial Pathogens
journal, August 2010


Global Transcriptome Analysis of Shewanella oneidensis MR-1 Exposed to Different Terminal Electron Acceptors
journal, September 2005


Identification of Genes Involved in Cytochrome c Biogenesis in Shewanella oneidensis, Using a Modified mariner Transposon
journal, August 2005


Transcriptomic and Proteomic Characterization of the Fur Modulon in the Metal-Reducing Bacterium Shewanella oneidensis
journal, December 2004


PBAD-Based Shuttle Vectors for Functional Analysis of Toxic and Highly Regulated Genes in Pseudomonas and Burkholderia spp. and Other Bacteria
journal, October 2008

  • Qiu, D.; Damron, F. H.; Mima, T.
  • Applied and Environmental Microbiology, Vol. 74, Issue 23
  • DOI: 10.1128/AEM.01369-08

GeoChip: a comprehensive microarray for investigating biogeochemical, ecological and environmental processes
journal, May 2007

  • He, Zhili; Gentry, Terry J.; Schadt, Christopher W.
  • The ISME Journal, Vol. 1, Issue 1
  • DOI: 10.1038/ismej.2007.2

Characterization of NaCl tolerance in Desulfovibrio vulgaris Hildenborough through experimental evolution
journal, April 2013


Genome-Wide Identification of Transcription Start Sites, Promoters and Transcription Factor Binding Sites in E. coli
journal, October 2009


An Ochrobactrum anthropi Gene Conferring Paraquat Resistance to the Heterologous Host Escherichia coli
journal, July 2001

  • Won, Sung-Hye; Lee, Byung-Hyun; Lee, Hyo-Shin
  • Biochemical and Biophysical Research Communications, Vol. 285, Issue 4
  • DOI: 10.1006/bbrc.2001.5268

An improved staining procedure for the detection of the peroxidase activity of cytochrome P-450 on sodium dodecyl sulfate polyacrylamide gels
journal, September 1976


Combined Genomics and Experimental Analyses of Respiratory Characteristics of Shewanella putrefaciens W3-18-1
journal, June 2013

  • Qiu, Dongru; Wei, Hehong; Tu, Qichao
  • Applied and Environmental Microbiology, Vol. 79, Issue 17
  • DOI: 10.1128/AEM.00619-13

Phototransformations of 5-Aminolevulinic Acid–induced Protoporphyrin IX in vitro: A Spectroscopic Study¶
journal, January 2000


Comparative systems biology across an evolutionary gradient within the Shewanella genus
journal, September 2009

  • Konstantinidis, Konstantinos T.; Serres, Margrethe H.; Romine, Margaret F.
  • Proceedings of the National Academy of Sciences, Vol. 106, Issue 37
  • DOI: 10.1073/pnas.0902000106

Pyruvate and Lactate Metabolism by Shewanella oneidensis MR-1 under Fermentation, Oxygen Limitation, and Fumarate Respiration Conditions
journal, September 2011

  • Pinchuk, Grigoriy E.; Geydebrekht, Oleg V.; Hill, Eric A.
  • Applied and Environmental Microbiology, Vol. 77, Issue 23
  • DOI: 10.1128/AEM.05382-11

Isolation and characterization of visible light-sensitive mutants of Escherichia coli K12
journal, June 1991

  • Miyamoto, Kazumasa; Nakahigashi, Kenji; Nishimura, Koichi
  • Journal of Molecular Biology, Vol. 219, Issue 3
  • DOI: 10.1016/0022-2836(91)90180-E

Impact of mutations in hemA and hemH genes on pyoverdine production by Pseudomonas fluorescens ATCC17400
journal, November 2001


Metal Binding to Saccharomyces cerevisiae Ferrochelatase
journal, November 2002

  • Karlberg, Tobias; Lecerof, David; Gora, Monika
  • Biochemistry, Vol. 41, Issue 46
  • DOI: 10.1021/bi0260785

Amino Acid Residues His183 and Glu264 in Bacillus subtilis Ferrochelatase Direct and Facilitate the Insertion of Metal Ion into Protoporphyrin IX ,
journal, January 2007

  • Hansson, Mattias D.; Karlberg, Tobias; Rahardja, Muhammad Arys
  • Biochemistry, Vol. 46, Issue 1
  • DOI: 10.1021/bi061760a

Human ferrochelatase is an iron-sulfur protein
journal, January 1994

  • Dailey, Harry A.; Finnegan, Michael G.; Johnson, Michael K.
  • Biochemistry, Vol. 33, Issue 2
  • DOI: 10.1021/bi00168a003

The extracytoplasmic function sigma factors: role in bacterial pathogenesis
journal, December 2004


Alternative Sigma Factors and Their Roles in Bacterial Virulence
journal, December 2005


Two Coregulated Efflux Transporters Modulate Intracellular Heme and Protoporphyrin IX Availability in Streptococcus agalactiae
journal, April 2010