The functional and optimal expression of genes is crucial
for survival of all living organisms. Numerous experiments
and efforts have been performed to reveal the mechanisms
required for the functional and optimal expression of human
genes. The yeast Saccharomyces cerevisiae has evolved
independently of humans for billions of years. Nevertheless,
S. cerevisiae has many conserved genes and expression mechanisms
that are similar to those in humans. Yeast is the most
commonly used model organism for studying the function
and expression mechanisms of human genes because it has
a relatively simple genome structure, which is easy to manipulate.
Many previous studies have focused on understanding
the functions and mechanisms of human proteins using
orthologous genes and biological systems of yeast. In this
review, we mainly introduce two recent studies that replaced
human genes and nucleosomes with those of yeast. Here, we
suggest that, although yeast is a relatively small eukaryotic
cell, its humanization is useful for the direct study of human
proteins. In addition, yeast can be used as a model organism
in a broader range of studies, including drug screening.
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A Humanized Yeast Model for Studying TRAPP Complex Mutations; Proof-of-Concept Using Variants from an Individual with a TRAPPC1-Associated Neurodevelopmental Syndrome Erta Zykaj, Chelsea Abboud, Paria Asadi, Simane Warsame, Hashem Almousa, Miroslav P. Milev, Brittany M. Greco, Marcos López-Sánchez, Drago Bratkovic, Aashiq H. Kachroo, Luis Alberto Pérez-Jurado, Michael Sacher Cells.2024; 13(17): 1457. CrossRef
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A nitrate-reducing Fe(II)-oxidizing bacterial strain, F8825T,
was isolated from the Fe(II)-rich sediment of an urban creek
in Pearl River Delta, China. The strain was Gram-negative,
facultative chemolithotrophic, facultative anaerobic, nonspore-
forming, and rod-shaped with a single flagellum. Phylogenetic
analysis based on 16S rRNA gene sequencing indicated
that it belongs to the genus Ciceribacter and is most
closely related to C. lividus MSSRFBL1T (99.4%), followed
by C. thiooxidans F43bT (98.8%) and C. azotifigens A.slu09T
(98.0%). Fatty acid, polar lipid, respiratory quinone, and
DNA G + C content analyses supported its classification in
the genus Ciceribacter. Multilocus sequence analysis of concatenated
16S rRNA, atpD, glnII, gyrB, recA, and thrC suggested
that the isolate was a novel species. DNA–DNA hybridization
and genome sequence comparisons (90.88 and
89.86%, for values of ANIm and ANIb between strains F8825T
with MSSRFBL1T, respectively) confirmed that strain F8825T
was a novel species, different from C. lividus MSSRFBL1T,
C. thiooxidans F43bT, and C. azotifigens A.slu09T. The physiological
and biochemical properties of the strain, such as
carbon source utilization, nitrate reduction, and ferrous ion
oxidation, further supported that this is a novel species. Based
on the polyphasic taxonomic results, strain F8825T was identified
as a novel species in the genus Ciceribacter, for which
the name Ciceribacter ferrooxidans sp. nov. is proposed.
The type strain is F8825T (= CCTCC AB 2018196T = KCTC
62948T).
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Strain KSNA2T, a Gram-negative, moderately halophilic, facultatively
anaerobic, motile, rod-shaped bacterium, was isolated
from the surface-sterilized stem tissue of a beach morning
glory (Calystegia soldanella) plant in Chuja Island, Jejudo,
Republic of Korea. Phylogenetic analysis based on 16S
rRNA gene and whole-genome sequences revealed that strain
KSNA2T formed a distinct lineage within the family Enterobacteriaceae,
with the highest 16S rRNA gene sequence similarity
to Izhakiella australiensis KCTC 72143T (96.2%) and
Izhakiella capsodis KCTC 72142T (96.0%), exhibited 95.5–
95.9% similarity to other genera in the family Enterobacteriaceae
and Erwiniaceae. Conserved signature indels analysis
elucidated that strain KSNA2T was delimited into family
Enterobacteriaceae. KSNA2T genome comprises a circular
chromosome of 5,182,800 bp with 56.1% G + C content. Digital
DNA-DNA relatedness levels between strain KSNA2T
and 18 closely related species were 19.3 to 21.1%. Average
nucleotide identity values were between 72.0 and 76.7%.
Growth of strain KSNA2T was observed at 4 to 45°C (optimum,
25°C) and pH 5.0 to 12.0 (optimum, pH 7.0) in the
presence of 0 to 11% (w/v) NaCl (optimum, 0–7%). The major
cellular fatty acids (> 10%) were C16:0 followed by summed
feature 8 (C18:1 ω7c and/or C18:1 ω6c), summed feature
3 (C16:1 ω7c and/or C16:1 ω6c), C17:0 cyclo, and C14:0. The major
isoprenoid quinone was ubiquinone-8 (Q-8). With combined
phylogenetic, genomic, phenotypic, and chemotaxonomic
features, strain KSNA2T represents a novel species of
a new genus in the family Enterobacteriaceae, for which the
name Jejubacter calystegiae gen. nov., sp. nov. is proposed.
The type strain is KSNA2T (= KCTC 72234T = CCTCC AB
2019098T).
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The gut microbiome of captive primates can provide a window
into their health and disease status. The diversity and
composition of gut microbiota are influenced by not only
host phylogeny, but also host diet. Old World monkeys (Cercopithecidae)
are divided into two subfamilies: Cercopithecinae
and Colobinae. The diet and physiological digestive features
differ between these two subfamilies. Accordingly, highthroughput
sequencing was used to examine gut microbiota
differences between these two subfamilies, using data from
29 Cercopithecinae individuals and 19 Colobinae individuals
raised in captivity. Through a comparative analysis of operational
taxonomic units (OTUs), significant differences in the
diversity and composition of gut microbiota were observed
between Cercopithecinae and Colobinae. In particular, the gut
microbiota of captive Old World monkeys clustered strongly
by the two subfamilies. The Colobinae microbial diversity was
higher than that of Cercopithecinae. Additionally, Firmicutes,
Lactobacillaceae, Veillonellaceae, and Prevotella abundance
were higher in Cercopithecinae, while Bacteroidetes, Ruminococcaceae,
Christensenellaceae, Bacteroidaceae, and Acidaminococcaceae
abundance were higher in Colobinae. PICRUSt
analysis revealed that the predicted metagenomes of metabolic
pathways associated with proteins, carbohydrates, and
amino acids were significantly higher in Colobinae. In the
context of host phylogeny, these differences between Cercopithecinae
and Colobinae could reflect adaptations associated
with their respective diets. This well-organized dataset is a
valuable resource for future related research on primates and
gut microbiota. Moreover, this study may provide useful insight
into animal management practices and primate conservation.
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The study of the human gut microbiome is essential in microbiology
and infectious diseases as specific alterations in the
gut microbiome might be associated with various pathologies,
such as chronic inflammatory disease, intestinal infection
and colorectal cancer. To identify such dysregulations,
several strategies are being used to create a repertoire of the
microorganisms composing the human gut microbiome. In
this study, we used the “microscomics” approach, which consists
of creating an ultrastructural repertoire of all the cell-like
objects composing stool samples from healthy donors using
transmission electron microscopy (TEM). We used TEM to
screen ultrathin sections of 8 resin-embedded stool samples.
After exploring hundreds of micrographs, we managed to
elaborate ultrastructural categories based on morphological
criteria or features. This approach explained many inconsistencies
observed with other techniques, such as metagenomics
and culturomics. We highlighted the value of our cultureindependent
approach by comparing our microscopic images
to those of cultured bacteria and those reported in the
literature. This study helped to detect “minimicrobes” Candidate
Phyla Radiation (CPR) for the first time in human
stool samples. This “microscomics” approach is non-exhaustive
but complements already existing approaches and adds
important data to the puzzle of the microbiota.
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Two bacterial strains designated NKC220-2T and NKC851-2
were isolated from commercial kimchi from different areas
in Korea. The strains were Gram-positive, aerobic, oxidaseand
catalase-positive, rod-shaped, spore-forming, non-motile,
and halophilic bacteria. Both strains grew without NaCl,
unlike type species in the genus Lentibacillus. The optimal
pH for growth was 8.0, higher than that of the type species
in the genus Lentibacillus, although growth was observed at
pH 5.5–9.0. 16S rRNA gene sequence-based phylogenetic analysis
indicated that the two strains (99.3–99.9% similarity)
are grouped within the genus Lentibacillus and most closely
related to Lentibacillus juripiscarius IS40-3T (97.4–97.6% similarity)
isolated from fish sauce in Thailand. OrthoANI value
between two novel strains and Lentibacillus lipolyticus SSKP1-
9T (79.5–79.6% similarity) was far lower than the species demarcation
threshold. Comparative genomic analysis displayed
differences between the two strains as well as among other
strains belonging to Lentibacillus. Furthermore, each isolate
had strain-specific groups of orthologous genes based on pangenome
analysis. Genomic G + C contents of strains NKC-
220-2T and NKC851-2 were 41.9 and 42.2 mol%, respectively.
The strains contained meso-diaminopimelic acid in their
cell walls, and the major menaquinone was menaquinone-7.
Phosphatidylglycerol, diphosphatidylglycerol, and an unidentified
glycolipid, aminophospholipid, and phospholipid were
the major polar lipid components of both strains. The major
cellular fatty acids of the strains were anteiso-C15:0 and anteiso-
C17:0. Based on phenotypic, genomic, phylogenetic, and
chemotaxonomic features, strains NKC220-2T and NKC851-2
represent novel species of the genus Lentibacillus, for which
the name Lentibacillus cibarius sp. nov. is proposed. The type
strain is NKC220-2T (= KACC 21232T = JCM 33390T).
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The widespread use of the organochlorine insecticide lindane
in the world has caused serious environmental problems.
The main purpose of this paper is to investigate the potency
of several Phlebia species of white rot fungi to degrade, transform
and mineralize lindane, and to provide the feasibility
of using white rot fungi for bioremediation at contaminated
sites. Based on tolerance experiment results, Phlebia brevispora
and Phlebia lindtneri had the highest tolerance to lindane
and were screened by degradation tests. After 25 days of
incubation, P. brevispora and P. lindtneri degraded 87.2 and
73.3% of lindane in low nitrogen medium and 75.8 and 64.9%
of lindane in high nitrogen medium, respectively. Several unreported
hydroxylation metabolites, including monohydroxylated,
dehydroxylated, and trihydroxylated products, were detected
and identified by GC/MS as metabolites of lindane.
More than 10% of [14C] lindane was mineralized to 14CO2 by
two fungi after 60 days of incubation, and the mineralization
was slightly promoted by the addition of glucose. Additionally,
the degradation of lindane and the formation of metabolites
were efficiently inhibited by piperonyl butoxide, demonstrating
that cytochrome P450 enzymes are involved in the fungal
transformation of lindane. The present study showed that
P. brevispora and P. lindtneri were efficient degraders of lindane;
hence, they can be applied in the bioremediation process
of lindane-contaminated sites.
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Neisseria gonorrhoeae, an obligatory human pathogen causes
the sexually transmitted disease gonorrhea, which remains
a global health problem. N. gonorrhoeae primarily infects the
mucosa of the genitourinary tract, which in women, is colonized
by natural microbiota, dominated by Lactobacillus spp.,
that protect human cells against pathogens. In this study, we
demonstrated that precolonization of human epithelial cells
with Lactobacillus crispatus, one of the most prevalent bacteria
in the female urogenital tract, or preincubation with the
L. crispatus enolase or glutamine synthetase impairs the adhesion
and invasiveness of N. gonorrhoeae toward epithelial
cells, two crucial steps in gonococcal pathogenesis. Furthermore,
decreased expression of genes encoding the proinflammatory
cytokines, TNFα and CCL20, which are secreted as
a consequence of N. gonorrhoeae infection, was observed in
N. gonorrhoeae-infected epithelial cells that had been precolonized
with L. crispatus or preincubated with enolase and
glutamine synthetase. Thus, our results indicate that the protection
of human cells against N. gonorrhoeae infection is a
complex process and that L. crispatus and its proteins enolase
and glutamine synthetase can have a potential role in protecting
epithelial cells against gonococcal infection. Therefore,
these results are important since disturbances of the microbiota
or of its proteins can result in dysbiosis, which is associated
with increased susceptibility of epithelium to pathogens.
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Carbapenems are a class of β-lactam antibiotics with a broad
antimicrobial activity spectrum. Owing to their sturdy structures
resistant to most β-lactamases, they have been regarded
as one of the last-resort antibiotics for combating multidrugresistant
bacterial infections. However, the emergence of carbapenem
resistance increases predominantly in nosocomial
pathogens. To prevent spread of carbapenem resistance in
early stages, it is imperative to develop rapid diagnostic tests
that will substantially reduce the time and cost in determining
carbapenem resistance. Thus, we devised a staining-based
diagnostic method applicable to three different Gram-negative
pathogens of Acinetobacter baumannii, Escherichia coli,
and Klebsiella pneumoniae, all with the high potential to develop
carbapenem resistance. Regardless of the resistance mechanisms
presented by bacterial species and strains, double
staining with propidium iodide (PI) and alamar blue (AB)
identified resistant bacteria with an average sensitivity of
95.35%, 7 h after imipenem treatments in 343 clinical isolates.
Among the three species tested, A. baumannii showed the
highest diagnostic sensitivity of 98.46%. The PI and ABmediated
staining method could be a promising diagnostic method with high-throughput efficacy and low cost.
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Enterovirus A71 (EV71), the main etiological agent of handfoot-
mouth disease (HFMD), circulates in many areas of the
world and has caused large epidemics since 1997, especially
in the Asia-Pacific region. In this study, we determined the
full-genome sequence of CMC718, a newly isolated EV71
strain in Korea. The CMC718 genome was 7,415 nucleotides
in length and was confirmed by whole-genome phylogenetic
analysis to belong to the B5 genotype. In particular, CMC718
demonstrated maximum identity with strain M988 of the B5
genotype and numerous amino acid variants were detected
in the 3D domain of the viral protein P3, which is consistent
with the mutation pattern of a B5 strain isolated in 2012–2013.
Comparison of the CMC718 sequence with other EV71 reference
strains confirmed the relationship and genetic variation
of CMC718. Our study was a full-genome sequence analysis
of the first EV71 strain of the B5 genotype isolated in
South Korea. This information will be a valuable reference
for the development of methods for the detection of recombinant
viruses, the tracking of infections, and the diagnosis
of EV71.