STARVATION AND STRINGENT STRESS

PRESENTED BY:
TEJASWINI C S ,
1ST MSc, 2ND SEMESTER,
MAHARANI’S SCIENCE COLLEGE
FOR WOMEN-MYSORE.
INDEX
➢ Starvation in microbes

➢ Starvation stress in E.coli

➢ Cellular changes in stationary phase

➢ Stringent response

➢ Stringent response in E.coli

➢ Conclusion
➢ Reference
 Starvation in microbes
➢ A significant environment impact on bacteria is stress, which in effecting a myriad of
adaptive and protective responses, alters gene expression patterns and cell physiology and
do influence antimicrobial susceptibility. This occurs indirectly, as a result of stress-
induced growth cessation or dormancy (due to starvation).

➢ In the lifecycle of microorganisms, prolonged starvation is prevalent and sustaining life

during starvation periods is a vital task.

➢ Starvation survival is defined as the ability to withstand long periods without energy
yielding substrates.

➢ Nutritional stress can change bacterial morphology.

➢ Many species of Gram-positive bacteria produce dormant spores in response to
starvation. By contrast, many Gram negative bacteria develop resistance cells without
dormancy.

➢ Starvation of many non differentiating bacteria such as E.coli and Salmonella respond to
starvation of an essential nutrient like carbon by inducing expression 50 or so new
proteins or preexisting proteins.

➢ During bacteria starvation the USP genes upregulated will often arrest cell growth and
promotes its metabolism to adapt nutrients .The universal stress protein (USP) domain
is a super family of conserved genes which is induced by many environmental stressors
such as nutrient starvation.

➢ FadR (regulator of fatty acid metabolism) binds to medium/long chain fatty acyl CoA
and repress fatty acid biosynthesis genes and activates fatty acid degradation genes.
➢ After Enter into stationary phase due to starvation and other strategy
-Morphological changes Ex: Endospore formation
- Decrease in size, protoplast shrinkage, and nucleoid condensation
- Rops Protein assists RNA polymerase in transcribe genes for starvation proteins

➢ Lrp (leucine responsive protein) which controls certain aspects of amino acids
metabolism.

➢ Low levels of carbon, nitrogen or phosphorus, as well as amino acid starvation , trigger
RpoS Synthesis. Induction of RpoS changes the gene expression pattern, aiming to
produce a more resistant cell.
➢ Low levels of carbon, nitrogen or phosphorus, as well as amino acid starvation , trigger
RpoS Synthesis. Induction of RpoS changes the gene expression pattern, aiming to
produce a more resistant cell.

➢ Proteins RelA and SpoT are both in charge of adjusting ppGpp concentration with in
the cell. ppGpp is key factor (unusual nucleotide found during bacterial stress starvation)
in bacterial physiology because it responds rapidly to diverse stresses, shutting down
growth and priming cellular defensive and adaptive processes
 Starvation stress in E.coli
➢ When microorganisms encounter nutrient deprivation or starvation conditions, they can
carry out starvation-induced activities such as production of degradative enzymes such as
proteases, lipases, and substrate capturing enzymes such as glutamine synthetase, and
alkaline phosphatase.

➢ When E.coli cells enter the stationary phase, a depletion of nutrients or starvation
conditions are encountered, and a number of morphological and physiological changes
occur.

➢ Cells became smaller and rounder, the cells accumulated storage compounds such as
glycogen and polyphosphate, and the DNA condensed.

➢ There were a number of changes in the fatty acid composition of their membrane and in
the protein composition of both inner and outer membranes in E.coli
➢ Rpos is the major regulator of the stationary phase or general stress response in E.coli and
other enteric bacteria. This alternative sigma factor has been recognized as a key factor in
producing greater resistance of stationary phase and stressed cells.

➢ In E.coli, two classes of genes encoding starvation proteins have been defined. These are
the cst genes, controlled by carbon starvation, and the pex genes, controlled by carbon,
nitrogen, or phosphorus starvation. The Pex (postexponential) proteins played a
significant role in coping with starvation stress in E.coli

➢ Several Pex proteins were induced during heat or oxidation stress and have been
correlated with the enhanced thermal and oxidative resistance that developed during
starvation in E.coli.
 Stringent response
➢ Stringent response, also called stringent control, is a stress response that occurs in
bacteria in reaction to amino-acid starvation, fatty acid limitation, heat shock, and other
stress conditions.

➢ During this control, bacteria experience conditions that limit the availability of one or
more amino acids or exhaust their primary carbon source, growth stops temporarily and
rapid adjustments in metabolism occur.

➢ Here cells respond to amino acid starvation by downregulating rRNA biosynthesis,

ribosomal proteins and DNA replication, and upregulating the levels of Rpos, stress
protein and amino acid biosynthesis.

➢ Stringent response collectively enhances cellular viability during periods of amino acid or
energy limitation and allows recovery and reinitiation of growth when conditions
improve.
➢ In bacteria stringent response is mediated by a variety of RelA/SpoT Homologue(RSH)
Proteins, Proteins RelA and SpoT are both in charge of adjusting ppGpp concentration
within the cell

➢ RelA and SpoT are both in charge of adjusting ppGpp concentration within the cell
- Chemical reaction catalyzed by RelA:
ATP + GTP AMP + ppGpp
- Chemical reaction catalyzed by SpoT:
ppGpp GDP + PPi or pppGpp
➢ The disabling of the stringent response by disruption of relA and spot in Pseudomonas
produced in infectious cells and biofilms characterized by nutrient limitation, causes
greater susceptibility to antibiotics.

➢ Translational GTPases involved in protein biosynthesis are also affected by ppGpp with
Initiation Factor 2 or IF2(controls the entry of tRNA onto ribosome) being the main target
 Stringent response in E.coli
➢ When bacteria grows in one or more than one amino acid deficiency, they shut down
their wide range of activities
➢ It is a mechanism of surviving hard times
➢ Bacteria performs only minimum activities
➢ Bacteria causes massive reduction in tRNA and rRNA levels
➢ Deprivation of single amino acid or mutation in Amino-acyl-tRNA-synthetase sufficient
to initiate stringent response
 What happens during stringent response?
➢ Several metabolic adjustments takes place
▪ Rate of protein degradation increases
▪ Synthesis of stress proteins and proteins
required for amino acid synthesis increases
▪ Stress(oxidative and osmotic stress)
survival genes are upregulated

▪ rRNA and tRNA synthesis decreases by 10 fold

▪ mRNA synthesis decreases by 3 fold
▪ Synthesis of carbohydrates, nucleotides and lipids also decreases
 Stringent response is controlled by 2 unusual nucleotides

1) ppGpp 2) pppGpp
-Guanosine tetraphosphate -Guanosine Penta phosphate
-diphosphate attached to both -triphosphate attached to both 5’
5’ and 3’positions. and diphosphate to 3’ position.

Also called as “Alarmone”

 How stringent response starts?
➢ Normally amino-acyl-tRNA binds to A-site of ribosome.

➢ But, when any amino acid is not there , the uncharged tRNA gain entry into A-site.

➢ Presence of uncharged tRNA at A-site triggers the stringent response

Relaxed mutants
➢ The bacterial mutants that cannot produce stringent response are called as relaxed
mutants.
➢ Most commonly mutation in gene relA that codes for stringent factor produces relaxed
mutants
 CONCLUSION
➢ Starvation survival is defined as the ability to withstand long periods without energy
yielding substrates
➢ Cells respond to amino acid starvation by downregulating rRNA biosynthesis, ribosomal
proteins and DNA replication, and upregulating the levels of Rpos, stress protein and
amino acid biosynthesis. This phenomenon is known as the stringent response
➢ In Gram-negative bacteria the starvation response triggers the alternative sigma factor
RpoS, which controls up to 10% of the E.coli genes
➢ The bacterial stress response allows bacteria to adapt to changing and unpleasant
conditions in their immediate environment. Distinct bacterial systems perceive different
environmental changes and respond accordingly. While the surrounding environment
commonly causes stress in microorganisms, the expansion of microbial cells can also
result in the production of stress such as hunger and acidity.
➢ The stringent response is mediated by (p) ppGpp and regulated by RelA and SpoT, which
affect antibiotic susceptibility.
 REFERENCE

https://pubmed.ncbi.nlm.nih.gov/30852584/

https://www.researchgate.net/publication/319199568_Starvation_exercise_and_the_stres
s_response

https://helioscrep.cssdm.gouv.qc.ca/p//record?ID=M52465s&readBook=Essential%20physi
ological%20biochemistry%20an%20organ%20based%20approach%20pdf.pdf
Thank you