1. Presented by: Sara Zapata Loaiza - Valentina David Martinez
Semestre III
Universidad Pontificia Bolivariana
Uropathogenic Escherichia Coli Virulence Factor α
Hemolysin Reduces Histone Acetylation to Inhibit
Expression of Pro-inflammatory Cytokine Genes
Zhengguo Zhang, Ming Wang, Yu Zhang, Yiming Zhang, Marek
Bartkuhn, Melanie Markmann, Hamid Hossain, Trinad Chakraborty,
Sandra B. Hake, Zhankui Jia, Andreas Meinhardt, Sudhanshu
Bhushan
2. Uropathogenic Escherichia coli
Escherichia coli is a
gram-negative,
anaerobic, rod-shaped,
coliform bacterium that
is commonly found in the
lower intestine of warm-
blooded organisms.
Symptoms
- Dysuria, hematuria, pyuria.
- Needing to pee more than normal.
- Feeling like you need to pee again
after going to the toilet.
- Dark urine, cloudy or strong-
smelling.
- Pelvic pain.
- Fever, chills, headache, fatigue.
Genetic and molecular biology
• The structure of its genome is highly flexible,
allows mobility of genetic material through
insertion sequences
Model organism in laboratories
• Due to its growth speed and few nutritional
requirements
3. α Hemolysin and Pro-inflammatory Cytokine Genes
Alpha hemolysin toxin (HlyA)
is an important virulence factor
produced by uropathogenic
strains of Escherichia coli and
is involved in various
pathologies such as meningitis,
septicemia, peritonitis or
diseases of the urinary tract.
Proinflammatory cytokines are a
category of small proteins
important in cell signaling. They
are responsible for intercellular
communication, functions of cell
proliferation and differentiation,
growth and modulation of
immunoglobulin secretion.
Cytokines involved in the inflammatory
process of the UPEC
IL-1β
IL-6
IL-8
4. Object
We aimed to establish how HlyA-mediated
inhibition of inflammatory gene expression was
achieved and whether epigenetic modifications
were involved in the process.
5. Methods
Bacterial strains and cell line
It’s used to diagnose infections, to test new
drugs, and for research.
For this study, it was to analyze which of the
strains that were used is more virulent.
6. Methods
Histopathology and hematoxylin and eosin
staining
Helps identify different types of cells and tissues,
and obtain important information about the
characteristics, shape, and cellular structure of a
tissue sample and it is used for the diagnosis of
diseases.
In this study was used to see if when doing acetate
treatment in mice with cystitis caused by UPEC, the
taken sections of bladder would change.
7. Methods
Immunoblot
A test used to evaluate the reaction between an
antigen and an antibody.
In this study immunoblot was done in order to find
the presence of the protein and virulent factor
alpha hemolysin in the samples.
8. Methods
Sodium dodecyl sulfate
polyacrylamide gel electrophoresis
(SDS-PAGE)
SDS-PAGE is an electrophoresis method
that allows protein separation by mass, to
analyze proteins.
It was used to determinate the acetylation
levels of histones when infected with UPEC
and HDM.
9. ResulTs
Immunoblot
- Beta actin = test control because it is a protein
present in all cell lines
- The longer the time, the expression of the
protein begins to decrease
10. ResulTs
Immunofluorescence
- The intensity of the fluorescence in UPEC and in
HDM increases compared to the control.
- UPEC and HDM were related to the expression of
the virulence of alpha hemolysin, that is why it
looks more fluorescent and means that there is
greater expression
11. ResulTs
SDS-PAGE
- The expression of H4K20me3 and H3K9me3 is
maintained over time = no knockdown ratio.
- With HDM, the band thickens over time = there is more
expression of those proteins
12. ResulTs
Immunoblot
- Inhibition of ACLY would indeed lead to
reduced histone acetylation.
- BMS-303141 treatment resulted in lower
H3 and H4 acetylation in a time- and
dose- dependent manner.
13. Discussion
AUTHOR STATEMENT RIGHT / WRONG
Zhang Z, et al. Recent investigation for
phatogen-mediated
reduction in histones
Wiles TJ, et al. HlyA regulates many
aspects of host cell
Pietrocola F, et al. Histone acetylation is
catalyzed by HAT
14. conclusions
Has allowed the advancement in
the knowledge of pathological
biological processes, which has
allowed the development of
treatments directed to specific
molecular pathways and more
accurate diagnosis
Provided the knowledge of a
complete sequence of human
DNA which constitutes a
powerful research tool that
has allowed the
advancement of the
development of new
therapies
Is constantly changing due to
the large amount of research
that is carried out being useful
to support different medical
activities such as diagnostic
methods
MOLECULAR BIOLOGY IN MEDICINE
