Role of aspartate ammonia-lyase in Pasteurella multocida.
Aspartate ammonia-lyase
Iron acquisition
Pasteurella multocida
Virulence
Journal
BMC microbiology
ISSN: 1471-2180
Titre abrégé: BMC Microbiol
Pays: England
ID NLM: 100966981
Informations de publication
Date de publication:
03 12 2020
03 12 2020
Historique:
received:
27
06
2020
accepted:
23
11
2020
entrez:
4
12
2020
pubmed:
5
12
2020
medline:
4
8
2021
Statut:
epublish
Résumé
Pasteurella multocida is responsible for a highly infectious and contagious disease in birds, leading to heavy economic losses in the chicken industry. However, the pathogenesis of this disease is poorly understood. We recently identified an aspartate ammonia-lyase (aspA) in P. multocida that was significantly upregulated under iron-restricted conditions, the protein of which could effectively protect chicken flocks against P. multocida. However, the functions of this gene remain unclear. In the present study, we constructed aspA mutant strain △aspA::kan and complementary strain C△aspA::kan to investigate the function of aspA in detail. Deletion of the aspA gene in P. multocida resulted in a significant reduction in bacterial growth in LB (Luria-Bertani) and MH (Mueller-Hinton) media, which was rescued by supplementation with 20 mM fumarate. The mutant strain △aspA::kan showed significantly growth defects in anaerobic conditions and acid medium, compared with the wild-type strain. Moreover, growth of △aspA::kan was more seriously impaired than that of the wild-type strain under iron-restricted conditions, and this growth recovered after supplementation with iron ions. AspA transcription was negatively regulated by iron conditions, as demonstrated by quantitative reverse transcription-polymerase chain reaction. Although competitive index assay showed the wild-type strain outcompetes the aspA mutant strain and △aspA::kan was significantly more efficient at producing biofilms than the wild-type strain, there was no significant difference in virulence between the mutant and the wild-type strains. These results demonstrate that aspA is required for bacterial growth in complex medium, and under anaerobic, acid, and iron-limited conditions.
Sections du résumé
BACKGROUND
Pasteurella multocida is responsible for a highly infectious and contagious disease in birds, leading to heavy economic losses in the chicken industry. However, the pathogenesis of this disease is poorly understood. We recently identified an aspartate ammonia-lyase (aspA) in P. multocida that was significantly upregulated under iron-restricted conditions, the protein of which could effectively protect chicken flocks against P. multocida. However, the functions of this gene remain unclear. In the present study, we constructed aspA mutant strain △aspA::kan and complementary strain C△aspA::kan to investigate the function of aspA in detail.
RESULT
Deletion of the aspA gene in P. multocida resulted in a significant reduction in bacterial growth in LB (Luria-Bertani) and MH (Mueller-Hinton) media, which was rescued by supplementation with 20 mM fumarate. The mutant strain △aspA::kan showed significantly growth defects in anaerobic conditions and acid medium, compared with the wild-type strain. Moreover, growth of △aspA::kan was more seriously impaired than that of the wild-type strain under iron-restricted conditions, and this growth recovered after supplementation with iron ions. AspA transcription was negatively regulated by iron conditions, as demonstrated by quantitative reverse transcription-polymerase chain reaction. Although competitive index assay showed the wild-type strain outcompetes the aspA mutant strain and △aspA::kan was significantly more efficient at producing biofilms than the wild-type strain, there was no significant difference in virulence between the mutant and the wild-type strains.
CONCLUSION
These results demonstrate that aspA is required for bacterial growth in complex medium, and under anaerobic, acid, and iron-limited conditions.
Identifiants
pubmed: 33272193
doi: 10.1186/s12866-020-02049-2
pii: 10.1186/s12866-020-02049-2
pmc: PMC7713322
doi:
Substances chimiques
Acids
0
Bacterial Proteins
0
Fumarates
0
Iron
E1UOL152H7
Aspartate Ammonia-Lyase
EC 4.3.1.1
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
369Subventions
Organisme : National Key Research and Development Program of China
ID : 2018YFD0500503
Organisme : National Natural Science Foundation of China
ID : 31873017
Organisme : Technical System of National Broiler Industry
ID : CARS-41-G13
Références
Mol Microbiol. 2008 Jul;69(1):77-93
pubmed: 18433445
J Appl Microbiol. 2013 Dec;115(6):1368-78
pubmed: 23957584
Appl Microbiol Biotechnol. 2019 May;103(10):4203-4215
pubmed: 30972460
Biochemistry. 1988 Dec 27;27(26):9089-93
pubmed: 2853974
Vet Microbiol. 2015 Aug 31;179(1-2):2-14
pubmed: 25888312
Front Microbiol. 2017 Jun 07;8:920
pubmed: 28638368
mBio. 2012 Oct 30;3(6):
pubmed: 23111869
Biochem J. 1994 Dec 1;304 ( Pt 2):321-31
pubmed: 7998964
Front Microbiol. 2018 Jun 06;9:1204
pubmed: 29928267
Vet Microbiol. 2003 Dec 30;97(3-4):191-9
pubmed: 14654290
Appl Environ Microbiol. 2019 Jan 23;85(3):
pubmed: 30446551
J Bacteriol. 1995 Nov;177(21):6255-62
pubmed: 7592392
Microb Pathog. 2019 Feb;127:257-266
pubmed: 30550841
J Bacteriol. 2000 Sep;182(17):4704-10
pubmed: 10940008
Appl Environ Microbiol. 2018 Oct 1;84(20):
pubmed: 30054363
BMC Genomics. 2007 Mar 13;8:72
pubmed: 17355629
J Gen Microbiol. 1976 Nov;97(1):73-82
pubmed: 792395
J Bacteriol. 1991 May;173(9):2879-87
pubmed: 2019560
PLoS One. 2015 Jul 13;10(7):e0132504
pubmed: 26168192
Infect Immun. 2005 Jan;73(1):226-34
pubmed: 15618158
Biochem J. 1986 Jul 15;237(2):547-57
pubmed: 3541901
Front Microbiol. 2015 Sep 04;6:926
pubmed: 26388863
FEBS Lett. 2010 Jun 3;584(11):2311-4
pubmed: 20371246
Avian Dis. 2017 Dec;61(4):491-502
pubmed: 29337619
Curr Top Microbiol Immunol. 2012;361:1-22
pubmed: 22643916
Vet Microbiol. 2017 Mar;201:162-169
pubmed: 28284604
3 Biotech. 2018 Jan;8(1):32
pubmed: 29291145
Gene. 1999 Oct 18;239(1):129-35
pubmed: 10571042
Infect Immun. 2005 Dec;73(12):8167-78
pubmed: 16299312
J Ind Microbiol Biotechnol. 2017 Mar;44(3):443-451
pubmed: 28120129
J Bacteriol. 2002 Jul;184(14):3965-74
pubmed: 12081969
FEMS Microbiol Lett. 2006 Dec;265(1):1-10
pubmed: 17107417
Microbiology (Reading). 2005 Jan;151(Pt 1):243-257
pubmed: 15632442
Avian Pathol. 2019 Jun;48(3):221-229
pubmed: 30640510
J Bacteriol. 2013 Nov;195(21):4854-64
pubmed: 23974032
Wei Sheng Wu Xue Bao. 2012 Apr 4;52(4):526-31
pubmed: 22799219
Trends Biotechnol. 1994 Mar;12(3):95-103
pubmed: 7764830
Clin Microbiol Rev. 1993 Apr;6(2):137-49
pubmed: 8472246
Sheng Wu Gong Cheng Xue Bao. 2017 Sep 25;33(9):1489-1512
pubmed: 28956396
J Microbiol. 2018 Jul;56(7):449-457
pubmed: 29948830
Vet Microbiol. 2013 Dec 27;167(3-4):713-8
pubmed: 24075356