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- Volume 58(7); July 2020
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Review
- [MINIREVIEW]Bacterial bug-out bags: outer membrane vesicles and their proteins and functions
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Kesavan Dineshkumar , Vasudevan Aparna , Liang Wu , Jie Wan , Mohamod Hamed Abdelaziz , Zhaoliang Su , Shengjun Wang , Huaxi Xu
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J. Microbiol. 2020;58(7):531-542. Published online June 10, 2020
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DOI: https://doi.org/10.1007/s12275-020-0026-3
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11
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Abstract
- Among the major bacterial secretions, outer membrane vesicles
(OMVs) are significant and highly functional. The proteins
and other biomolecules identified within OMVs provide
new insights into the possible functions of OMVs in bacteria.
OMVs are rich in proteins, nucleic acids, toxins and
virulence factors that play a critical role in bacteria-host interactions.
In this review, we discuss some proteins with multifunctional
features from bacterial OMVs and their role
involving the mechanisms of bacterial survival and defence.
Proteins with moonlighting activities in OMVs are discussed
based on their functions in bacteria. OMVs harbour many
other proteins that are important, such as proteins involved
in virulence, defence, and competition. Overall, OMVs are a
power-packed aid for bacteria, harbouring many defensive
and moonlighting proteins and acting as a survival kit in
case
of an emergency or as a defence weapon. In summary,
OMVs can be defined as bug-out bags for bacterial defence
and, therefore, survival.
Journal Articles
- Azohydromonas aeria sp. nov., isolated from air
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Han Xue , Chun-gen Piao , Dan-ran Bian , Min-wei Guo , Yong Li
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J. Microbiol. 2020;58(7):543-549. Published online June 27, 2020
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DOI: https://doi.org/10.1007/s12275-020-9423-x
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4
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Abstract
- A grey pink colored bacterium, strain t3-1-3T, was isolated
from the air at the foot of the Xiangshan Mountain in Beijing,
China. The cells are aerobic, Gram-stain-negative, non-sporeforming,
motile and coccoid-rod shaped (0.9–1.2 × 1.9–2.1
μm). Strain t3-1-3T was catalase-positive and oxidase-negative
and this strain grew at 4–42°C (optimum 28°C), a pH
of 4.0–9.0 (optimum pH 7.0) and under 0–2% (w/v) NaCl
(optimum 0–1% NaCl). A phylogenetic analysis based on 16S
rRNA gene sequences revealed that strain t3-1-3T was closely
related to Azohydromonas riparia UCM-11T (97.4% similarity),
followed by Azohydromonas australica G1-2T (96.8%)
and Azohydromonas ureilytica UCM-80T (96.7%). The genome
of strain t3-1-3T contains 6,895 predicted protein-encoding
genes, 8 rRNA genes, 62 tRNA genes and one sRNA
gene, as well as five potential biosynthetic gene clusters, including
clusters of genes coding for non-ribosomal peptide
synthetase (NRPS), bacteriocin and arylpolyene and two clusters
of genes for terpene. The predominant cellular fatty acids
(> 10.0% of the total) in strain t3-1-3T were summed feature
3 (C16:1ω7c and/or C16:1ω6c, 37.8%), summed feature 8
(C18:1ω7c and/or C18:1ω6c, 29.7%) and C16:0 (17.3%). Strain
t3-1-3T contained ubiquinone-8 (Q-8) as the predominant
respiratory quinone. The polar lipids of strain t3-1-3T comprised
phosphatidyl ethanolamine (PE), phosphatidyl glycerol
(PG), diphosphatidyl glycerol (DPG), an unidentified
glycolipid (GL), an unidentified aminophospholipid (APL),
two unidentified phospholipid (PL1-2) and five unidentified
lipid (L1-5). The DNA G + C content of the type strain
is 70.3%. The broader range of growth temperature, assimilation
of malic acid and trisodium citrate, presence of C18:3ω6c
and an unidentified glycolipid and absence of C12:0 2-OH and
C16:0iso differentiate strain t3-1-3T from related species. Based
on the taxonomic data presented in this study, we suggest
that strain t3-1-3T represents a novel species within the genus
Azohydromonas, for which the name Azohydromonas
aeria sp. nov. is proposed. The type strain of Azohydromonas
aeria is t3-1-3T (= CFCC 13393T = LMG 30135T).
- Burkholderia thailandensis outer membrane vesicles exert antimicrobial activity against drug-resistant and competitor microbial species
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Yihui Wang , Joseph P. Hoffmann , Chau-Wen Chou , Kerstin Höner zu Bentrup , Joseph A. Fuselier , Jacob P. Bitoun , William C. Wimley , Lisa A. Morici
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J. Microbiol. 2020;58(7):550-562. Published online April 11, 2020
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DOI: https://doi.org/10.1007/s12275-020-0028-1
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37
Citations
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Abstract
- Gram-negative bacteria secrete outer membrane vesicles
(OMVs) that play critical roles in intraspecies, interspecies,
and bacteria-environment interactions. Some OMVs, such
as those produced by Pseudomonas aeruginosa, have previously
been shown to possess antimicrobial activity against
competitor species. In the current study, we demonstrate that
OMVs from Burkholderia thailandensis inhibit the growth
of drug-sensitive and drug-resistant bacteria and fungi. We
show that a number of antimicrobial compounds, including
peptidoglycan hydrolases, 4-hydroxy-3-methyl-2-(2-nonenyl)-
quinoline (HMNQ) and long-chain rhamnolipid are
present in or tightly associate with B. thailandensis OMVs.
Furthermore, we demonstrate that HMNQ and rhamnolipid
possess antimicrobial and antibiofilm properties against methicillin-
resistant Staphylococcus aureus (MRSA). These findings
indicate that B. thailandensis secretes antimicrobial OMVs
that may impart a survival advantage by eliminating competition.
In addition, bacterial OMVs may represent an untapped
resource of novel therapeutics effective against biofilm-
forming and multidrug-resistant organisms.
- Long-term continuously monocropped peanut significantly disturbed the balance of soil fungal communities
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Mingna Chen , Jiancheng Zhang , Hu Liu , Mian Wang , LiJuan Pan , Na Chen , Tong Wang , Yu Jing , Xiaoyuan Chi , Binghai Du
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J. Microbiol. 2020;58(7):563-573. Published online April 22, 2020
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DOI: https://doi.org/10.1007/s12275-020-9573-x
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12
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Abstract
- Balancing soil microbial diversity and abundance is critical
to sustaining soil health, and understanding the dynamics of
soil microbes in a monocropping system can help determine
how continuous monocropping practices induce soil sickness
mediated by microorganisms. This study used previously
constructed gradient continuous monocropping plots and
four varieties with different monocropping responses were
investigated. The feedback responses of their soil fungal communities
to short-term and long-term continuous monocropping
were tracked using high-throughput sequencing techniques.
The analyses indicated that soil samples from 1 and
2 year monocropped plots were grouped into one class, and
samples from the 11 and 12 year plots were grouped into another,
regardless of variety. At the species level, the F. solani,
Fusarium oxysporum, Neocosmospora striata, Acrophialophora
levis, Aspergillus niger, Aspergillus corrugatus, Thielavia
hyrcaniae, Emericellopsis minima, and Scedosporium
aurantiacum taxa showed significantly increased abundances
in the long-term monocropping libraries compared to
the short-term cropping libraries. In contrast, Talaromyces
flavus, Talaromyces purpureogenus, Mortierella alpina, Paranamyces
uniporus, and Volutella citrinella decreased in
the long-term monocropping libraries compared to the shortterm
libraries. This study, combined with our previous study,
showed that fungal community structure was significantly
affected by the length of the monocropping period, but peanut
variety and growth stages were less important. The increase
in pathogen abundances and the decrease in beneficial
fungi abundances seem to be the main cause for the yield decline
and poor growth of long-term monocultured peanut.
Simplification of fungal community diversity could also contribute
to peanut soil sickness under long-term monocropping.
Additionally, the different responses of peanut varieties
to monocropping may be related to variations in their
microbial community structure.
- The putative C2H2 transcription factor RocA is a novel regulator of development and secondary metabolism in Aspergillus nidulans
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Dong Chan Won , Yong Jin Kim , Da Hye Kim , Hee-Moon Park , Pil Jae Maeng
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J. Microbiol. 2020;58(7):574-587. Published online April 22, 2020
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DOI: https://doi.org/10.1007/s12275-020-0083-7
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Abstract
- Multiple transcriptional regulators play important roles in
the coordination of developmental processes, including asexual
and sexual development, and secondary metabolism in the
filamentous fungus Aspergillus nidulans. In the present study,
we characterized a novel putative C2H2-type transcription
factor (TF), RocA, in relation to development and secondary
metabolism. Deletion of rocA increased conidiation and caused
defective sexual development. In contrast, the overexpression
of rocA exerted opposite effects on both phenotypes. Additionally,
nullifying rocA resulted in enhanced brlA expression
and reduced nsdC expression, whereas its overexpression
exerted the opposite effects. These results suggest that RocA
functions as a negative regulator of asexual development by
repressing the expression of brlA encoding a key asexual development
activator, but as a positive regulator of sexual development
by enhancing the expression of nsdC encoding a
pivotal sexual development activator. Deletion of rocA increased
the production of sterigmatocystin (ST), as well as the
expression of its biosynthetic genes, aflR and stcU. Additionally,
the expression of the biosynthetic genes for penicillin
(PN), ipnA and acvA, and for terrequinone (TQ), tdiB and
tdiE, was increased by rocA deletion. Thus, it appears that
RocA functions as a negative transcriptional modulator of the
secondary metabolic genes involved in ST, PN, and TQ biosynthesis.
Taken together, we propose that RocA is a novel
transcriptional regulator that may act either positively or negatively
at multiple target genes necessary for asexual and
sexual development and secondary metabolism.
- Melatonin attenuates microbiota dysbiosis of jejunum in short-term sleep deprived mice
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Ting Gao , Zixu Wang , Jing Cao , Yulan Dong , Yaoxing Chen
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J. Microbiol. 2020;58(7):588-597. Published online May 18, 2020
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DOI: https://doi.org/10.1007/s12275-020-0094-4
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27
Citations
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Abstract
- Our study demonstrated that sleep deprivation resulted in
homeostasis disorder of colon. Our study goes deeper into the
positive effects of melatonin on small intestinal microbiota
disorder caused by sleep deprivation. We successfully established
a multiplatform 72 h sleep deprivation mouse model
with or without melatonin supplementation, and analyzed the
change of small intestinal microbiota using high-throughput
sequencing of the 16S rRNA. We found melatonin supplementation
suppressed the decrease of plasma melatonin level
in sleep deprivation mice. Meanwhile, melatonin supplementation
improved significantly the reduction in OTU numbers
and the diversity and richness of jejunal microbiota and
the abundance of Bacteroidaeae and Prevotellaceae, as well
as an increase in the Firmicutes-to-Bacteroidetes ratio and
the content of Moraxellaceae and Aeromonadaceae in the jejunum
of sleep deprived-mice. Moreover, melatonin supplementation
reversed the change of metabolic pathway in sleep
deprived-mice, including metabolism, signal transduction
mechanisms and transcription etc, which were related to intestinal
health. Furthermore, melatonin supplementation inverted
the sleep deprivation-induced a decline of anti-inflammatory
cytokines (IL-22) and an increase of the ROS and proinflammatory
cytokines (IL-17) in jejunum. These findings
suggested that melatonin, similar to a probiotics agent, can
reverse sleep deprivation-induced small intestinal microbiota
disorder by suppressing oxidative stress and inflammation
response.
- Phenotypic characterization of a conserved inner membrane protein YhcB in Escherichia coli
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Chul Gi Sung , Umji Choi , Chang-Ro Lee
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J. Microbiol. 2020;58(7):598-605. Published online April 22, 2020
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DOI: https://doi.org/10.1007/s12275-020-0078-4
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8
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Abstract
- Although bacteria have diverse membrane proteins, the function
of many of them remains unknown or uncertain even
in Escherichia coli. In this study, to investigate the function
of hypothetical membrane proteins, genome-wide analysis
of phenotypes of hypothetical membrane proteins was performed
under various envelope stresses. Several genes responsible
for adaptation to envelope stresses were identified.
Among them, deletion of YhcB, a conserved inner membrane
protein of unknown function, caused high sensitivities to various
envelope stresses and increased membrane permeability,
and caused growth defect under normal growth conditions.
Furthermore, yhcB deletion resulted in morphological
aberration, such as branched shape, and cell division defects,
such as filamentous growth and the generation of chromosome-
less cells. The analysis of antibiotic susceptibility
showed that the yhcB mutant was highly susceptible to various
anti-folate antibiotics. Notably, all phenotypes of the yhcB
mutant were completely or significantly restored by YhcB
without the transmembrane domain, indicating that the localization
of YhcB on the inner membrane is dispensable for its
function. Taken together, our results demonstrate that YhcB
is involved in cell morphology and cell division in a membrane
localization-independent manner.
- Structural and sequence comparisons of bacterial enoyl-CoA isomerase and enoyl-CoA hydratase
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Jisub Hwang , Chang-Sook Jeong , Chang Woo Lee , Seung Chul Shin , Han-Woo Kim , Sung Gu Lee , Ui Joung Youn , Chang Sup Lee , Tae-Jin Oh , Hak Jun Kim , Hyun Park , Hyun Ho Park , Jun Hyuck Lee
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J. Microbiol. 2020;58(7):606-613. Published online April 22, 2020
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DOI: https://doi.org/10.1007/s12275-020-0089-1
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Abstract
- Crystal structures of enoyl-coenzyme A (CoA) isomerase from
Bosea sp. PAMC 26642 (BoECI) and enoyl-CoA hydratase
from Hymenobacter sp. PAMC 26628 (HyECH) were determined
at 2.35 and 2.70 Å resolution, respectively. BoECI
and HyECH are members of the crotonase superfamily and
are enzymes known to be involved in fatty acid degradation.
Structurally, these enzymes are highly similar except for the
orientation of their C-terminal helix domain. Analytical ultracentrifugation
was performed to determine the oligomerization
states of BoECI and HyECH revealing they exist as
trimers in solution. However, their putative ligand-binding
sites and active site residue compositions are dissimilar. Comparative
sequence and structural analysis revealed that the
active site of BoECI had one glutamate residue (Glu135), this
site is occupied by an aspartate in some ECIs, and the active
sites of HyECH had two highly conserved glutamate residues
(Glu118 and Glu138). Moreover, HyECH possesses a salt
bridge interaction between Glu98 and Arg152 near the active
site. This interaction may allow the catalytic Glu118 residue
to have a specific conformation for the ECH enzyme reaction.
This salt bridge interaction is highly conserved in known
bacterial ECH structures and ECI enzymes do not have this
type of interaction. Collectively, our comparative sequential
and structural studies have provided useful information to
distinguish and classify two similar bacterial crotonase superfamily
enzymes.
- Endophytic bacterial and fungal microbiota in different cultivars of cassava (Manihot esculenta Crantz)
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Hong Li , Chengliang Yan , Yanqiong Tang , Xiang Ma , Yinhua Chen , Songbi Chen , Min Lin , Zhu Liu
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J. Microbiol. 2020;58(7):614-623. Published online May 18, 2020
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DOI: https://doi.org/10.1007/s12275-020-9565-x
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Abstract
- Endophytes colonize tissues of healthy host plants and play
a crucial role in plant growth and development. However,
little attention has been paid to the endophytes of tuber crops
such as cassava, which is used as a staple food by approximately
800 million people worldwide. This study aimed to
elucidate the diversity and composition of endophytic bacterial
and fungal communities in different cassava cultivars
using high-throughput sequencing. Although no significant
differences in richness or diversity were observed among the
different cassava cultivars, the community compositions were
diverse. Two cultivars (SC124 and SC205) tolerant to root rot
exhibited similar community compositions, while two other
cultivars (SC10 and SC5), which are moderately and highly
susceptible to root rot, respectively, harboured similar community
compositions. Proteobacteria, Firmicutes, and Ascomycota
dominated the endophyte assemblages, with Weissella,
Serratia, Lasiodiplodia, Fusarium, and Diaporthe being the
predominant genera. The differentially abundant taxonomic
clades between the tolerant and susceptible cultivars were
mainly rare taxa, such as Lachnoclostridium_5, Rhizobium,
Lampropedia, and Stenotrophomonas. These seemed to be key
genera that affected the susceptibility of cassava to root rot.
Moreover, the comparison of KEGG functional profiles revealed
that ‘Environmental adaptation’ category was significantly
enriched in the tolerant cultivars, while ‘Infectious
diseases: Parasitic’ category was significantly enriched in the
susceptible cultivars. The present findings open opportunities
for further studies on the roles of endophytes in the susceptibility
of plants to diseases.
- Phosphorylation of tegument protein pp28 contributes to trafficking to the assembly compartment in human cytomegalovirus infection
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Jun-Young Seo , Jin Ah Heo , William J. Britt
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J. Microbiol. 2020;58(7):624-631. Published online June 27, 2020
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DOI: https://doi.org/10.1007/s12275-020-0263-5
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Abstract
- Human cytomegalovirus (HCMV) UL99 encodes a late tegument
protein pp28 that is essential for envelopment and
production of infectious virus. This protein is localized to
the endoplasmic reticulum-Golgi intermediate compartment
(ERGIC) in transfected cells but it localizes to the cytoplasmic
assembly compartment (AC) in HCMV-infected cells. Trafficking
of pp28 to the AC is required for the assembly of infectious
virus. The N-terminal domain (aa 1-61) of pp28 is
sufficient for trafficking and function of the wild type protein
during viral infection. However, residues required for
authentic pp28 trafficking with the exception of the acidic
cluster in the N-terminal domain of pp28 remain undefined.
Monitoring protein migration on SDS-PAGE, we found that
pp28 is phosphorylated in the virus-infected cells and dephosphorylated
in the viral particles. By generating substitution
mutants of pp28, we showed that three serine residues
(aa 41–43) and a tyrosine residue (aa 34) account for its phosphorylation.
The mutant forms of pp28 were localized to the
plasma membrane as well as the ERGIC in transfected cells.
Likewise, these mutant proteins were localized to the plasma
membrane as well as the AC in virus-infected cells. These results
suggested that phosphorylation of pp28 contributes to
its intracellular trafficking and efficient viral assembly and
incorporation.
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