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Abiotic stresses, such as heat, can drive protein misfolding and aggregation, leading to inhibition of cellular function and ultimately cell death. Unexpectedly, a thermotolerant Escherichia coli was identified from a pool of antibiotic resistant RNA polymerase β subunit (rpoB) mutants. This stress tolerant phenotype was characterized through exposure to high

Abiotic stresses, such as heat, can drive protein misfolding and aggregation, leading to inhibition of cellular function and ultimately cell death. Unexpectedly, a thermotolerant Escherichia coli was identified from a pool of antibiotic resistant RNA polymerase β subunit (rpoB) mutants. This stress tolerant phenotype was characterized through exposure to high temperature and ethanol. After 30-minute exposure of cells to 55°C or 25% ethanol, the mutant displayed 100 times greater viability than the wild-type, indicating that the rpoB mutation may have broadly affected the cellular environment to reduce protein misfolding and/or prevent protein aggregation. To further test this hypothesis, we examined thermotolerance of cells lacking heat shock chaperone DnaJ (Hsp40), which is a cochaperone of one of the most abundant and conserved chaperones, DnaK (Hsp70). The deletion of dnaJ led to severe growth defects in the wild-type, namely a slower growth rate and extreme filamentation at 42°C. The severity of the growth defects increased after additionally deleting DnaJ analog, CbpA. However, these defects were significantly ameliorated by the rpoB mutation. Finally, the rpoB mutant was found to be minimally affected by the simultaneous depletion of DnaK and DnaJ compared to the wild-type, which failed to form single colonies at 37°C and 42°C. Based on these observations, it is proposed that the rpoB mutant’s robust thermotolerant phenotype results from a cellular environment protective against protein aggregation or improper folding. The folding environment of the rpoB mutants should be further examined to elucidate the mechanism by which both antibiotic resistance and thermotolerance can be conferred.
ContributorsYeh, Melody (Author) / Misra, Rajeev (Thesis director) / Wang, Xuan (Committee member) / Kelly, Keilen (Committee member) / School of Life Sciences (Contributor) / School of International Letters and Cultures (Contributor) / School of Human Evolution & Social Change (Contributor) / Barrett, The Honors College (Contributor)
Created2020-05
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Description

Bacteria of the genus Wolbachia are
bacteria that live within the cells of their hosts. They infect a
wide range of arthropods (insects, arachnids, and crustaceans) and
some nematodes (parasitic roundworms). Scientists estimate that
Wolbachia exist in between seventeen percent and seventy-six percent of
arthropods

Bacteria of the genus Wolbachia are
bacteria that live within the cells of their hosts. They infect a
wide range of arthropods (insects, arachnids, and crustaceans) and
some nematodes (parasitic roundworms). Scientists estimate that
Wolbachia exist in between seventeen percent and seventy-six percent of
arthropods and nematodes. The frequency of the bacteria makes them
one of the most widespread parasites. In general, they are divided
into five groups, from A to E, depending of the species of their
host. They cause diverse reproductive and developmental changes on
their numerous invertebrate hosts. Several mechanisms, like the
feminization of the embryo's sexual characters, are involved in
those processes. To reproduce, Wolbachia often exploit their hosts'
reproductive processes. Additionally, they are symbiotic in that they are
necessary for the normal development of organisms in some species

Created2015-01-29
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Description

Between 1957 and 1959, Arthur Pardee, Francois Jacob, and Jacques Monod conducted a set of experiments at the Pasteur Institute in Paris, France, that was later called the PaJaMa Experiments, a moniker derived from the researchers' last names. In these experiments, they described how genes of a species of single-celled

Between 1957 and 1959, Arthur Pardee, Francois Jacob, and Jacques Monod conducted a set of experiments at the Pasteur Institute in Paris, France, that was later called the PaJaMa Experiments, a moniker derived from the researchers' last names. In these experiments, they described how genes of a species of single-celled bacteria, called Escherichia coli (E. coli), controlled the processes by which enzymes were produced in those bacteria. In 1959, the researchers published their results in a paper titled 'The Genetic Control and Cytoplasmic Expression of 'Inducibility' in the Synthesis of b-galactosidase by E. coli'. When they compared mutated strains of E. coli to a normal strain, Pardee, Jacob, and Monod identified the abnormal regulation processes and enzymes produced by the mutated genes. The results showed how enzymes break down the molecules that the bacteria ingested. The PaJaMas experiments uncovered some of the molecular mechanisms that regulate how some genes yield enzymes in many species.

Created2015-05-28
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Description

Lysogenic bacteria, or virus-infected bacteria, were the primary experimental models used by scientists working in the laboratories of the Pasteur Institute in Paris, France, during the 1950s and 1960s. Historians of science have noted that the use of lysogenic bacteria as a model in microbiological research influenced the scientific achievements

Lysogenic bacteria, or virus-infected bacteria, were the primary experimental models used by scientists working in the laboratories of the Pasteur Institute in Paris, France, during the 1950s and 1960s. Historians of science have noted that the use of lysogenic bacteria as a model in microbiological research influenced the scientific achievements of the Pasteur Institute's scientists. Francois Jacob and Jacques Monod used lysogenic bacteria to develop their operon model of gene regulation, to investigate the cellular regulatory mechanisms of the lysogenic life cycle, and to infer the process of cellular differentiation in the development of more complex eukaryotes.

Created2014-10-10
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Description

Leonard Colebrook was a physician who researched bacteria and infections in England during the twentieth century. In 1936, Colebrook deployed the antibiotic Prontosil to treat puerperal fever, a disorder that results from bacterial infections in the uterine tracts of women after childbirth or abortions. Colebrook also advanced care for burn

Leonard Colebrook was a physician who researched bacteria and infections in England during the twentieth century. In 1936, Colebrook deployed the antibiotic Prontosil to treat puerperal fever, a disorder that results from bacterial infections in the uterine tracts of women after childbirth or abortions. Colebrook also advanced care for burn patients by advocating for the creation of burn units in hospitals and by using antisepsis medication for burn wound infections. Colebrook’s work on treatments for puerperal fever reduced cases of puerperal fever throughout the world.

Created2017-05-25
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Description

In 2007, Philippe Horvath and his colleagues explained how bacteria protect themselves against viruses at Danisco, a Danish food company, in Dangé-Saint-Romain, France. Horvath and his team worked to improve the lifespan of bacteria cultures for manufacturing yogurt and ice cream. Specifically, they focused on bacteria’s resistance to bacteriophages, or

In 2007, Philippe Horvath and his colleagues explained how bacteria protect themselves against viruses at Danisco, a Danish food company, in Dangé-Saint-Romain, France. Horvath and his team worked to improve the lifespan of bacteria cultures for manufacturing yogurt and ice cream. Specifically, they focused on bacteria’s resistance to bacteriophages, or viruses that infect bacteria. Horvath and his colleagues found that the bacteria used to culture yogurt, Streptococcus thermophilus, has an adaptive immune system that can target specific viruses that have previously infected the bacteria. The immune system is called the CRISPR/cas system, or the clustered regularly interspaced short palindromic repeats/CRISPR associated protein system. Horvath and his colleagues explained how bacteria use CRISPR/cas as an immune system to target viruses and protect themselves from infection. The discovery informed the development of CRISPR/cas as a gene editing tool to modify bacterial, animal, and human genomes.

Created2017-11-15
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Description

In a series of experiments during mid 1930s, a team of researchers in New York helped establish that bacteria of the species Toxoplasma gondii can infect humans, and in infants can cause toxoplasmosis, a disease that inflames brains, lungs, and hearts, and that can organisms that have it. The team

In a series of experiments during mid 1930s, a team of researchers in New York helped establish that bacteria of the species Toxoplasma gondii can infect humans, and in infants can cause toxoplasmosis, a disease that inflames brains, lungs, and hearts, and that can organisms that have it. The team included Abner Wolf, David Cowen, and Beryl Paige. They published the results of their experiment in Human Toxoplasmosis: Occurrence in Infants as an Encephalomyelitis Verification of Transmission to Animals. Toxoplasmosis is an infection that causes inflammations in the brain (encephalitis), heart (myocarditis), and lungs (pneumonitis). The disease is caused in organisms that consume items contaminated by the protozoan parasite Toxoplasma gondii. The bacteria can transfer from pregnant women to their fetuses during pregnancy (congenitally), and it can lead those fetuses to develop physical deformities and mental disabilities. The 1930s experiments established Toxoplasma gondii as a human pathogen and helped increase research into congenital toxoplasmosis, enabling later researchers to develop measures to prevent against the disease in pregnant women.

Created2015-06-11
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Description

L'Institut Pasteur (The Pasteur Institute) is a non-profit private research institution founded by Louis Pasteur on 4 June 1887 in Paris, France. The Institute's research focuses on the study of infectious diseases, micro-organisms, viruses, and vaccines. As of 2014, ten scientists have received Nobel Prizes in physiology or medicine

L'Institut Pasteur (The Pasteur Institute) is a non-profit private research institution founded by Louis Pasteur on 4 June 1887 in Paris, France. The Institute's research focuses on the study of infectious diseases, micro-organisms, viruses, and vaccines. As of 2014, ten scientists have received Nobel Prizes in physiology or medicine for the research they have done at the Pasteur Institute. Contrary to the way genetics was studied in US research universities during the mid-twentieth century, the genetic research conducted at the Pasteur Institute at the same time did not rest on a conceptual separation between embryology and evolution. According to historian Michel Morange from the Ecole Normale Superieure in Paris, France, this difference enabled Pasteurian scientists to develop the concepts of regulatory genes and of developmental genes.

Created2014-08-19
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Description

Francois Jacob studied in
bacteria and bacteriophages at the Institut Pasteur in Paris, France,
in the second half of the twentieth century. In 1965, Jacob won the
Nobel Prize in Physiology or Medicine with Andre M. Lwoff and
Jacques L. Monod for their work on the

Francois Jacob studied in
bacteria and bacteriophages at the Institut Pasteur in Paris, France,
in the second half of the twentieth century. In 1965, Jacob won the
Nobel Prize in Physiology or Medicine with Andre M. Lwoff and
Jacques L. Monod for their work on the genetic control of enzyme
synthesis. Jacob studied how genes control and regulate metabolic
enzymes in the bacterium Escherichia
coli (E. coli) and in lysogenic
bacterial systems. He contributed to theories of transcriptional gene
regulation, the operon model, and the distinction between structural
and regulatory genes. Jacob also introduced the concept of
bricolage (tinkering) in evolutionary biology.

Created2014-09-29
Description

One of the identified health risk areas for human spaceflight is infectious disease, particularly involving environmental microorganisms already found on the International Space Station (ISS). In particular, bacteria belonging to the Burkholderia cepacia complex (Bcc) which can cause human disease in those who are immunocompromised, have been identified in the

One of the identified health risk areas for human spaceflight is infectious disease, particularly involving environmental microorganisms already found on the International Space Station (ISS). In particular, bacteria belonging to the Burkholderia cepacia complex (Bcc) which can cause human disease in those who are immunocompromised, have been identified in the ISS water supply. This present study characterized the effect of spaceflight analog culture conditions on Bcc to certain physiological stresses (acid and thermal as well as intracellular survival in U927 human macrophage cells). The NASA-designed Rotating Wall Vessel (RWV) bioreactor was used as the spaceflight analogue culture system in these studies to grow Bcc bacterial cells under Low Shear Modeled Microgravity (LSMMG) conditions. Results show that LSMMG culture increased the resistance of Bcc to both acid and thermal stressors, but did not alter phagocytic uptake in 2-D monolayers of human monocytes.

ContributorsVu, Christian-Alexander (Author) / Nickerson, Cheryl (Thesis director) / Barrila, Jennifer (Committee member) / Ott, Mark (Committee member) / Barrett, The Honors College (Contributor) / Harrington Bioengineering Program (Contributor)
Created2023-05