SRI SATHYA SAI INSTITUTE OF HIGHER LEARNING

(Deemed to be University)
Prashanthi Nilayam

Glutamine and glutamate modulate mutant Huntingtin aggregation in

Yeast Model of Huntington’s Disease
Neeraj Kalyan.G, Shakthi thangavel, Rajath D, Sai Sanwid Pradhan, Venketesh.S*

INTRODUCTION: TARGETED PATHWAY:

• Huntington’s disease is an autosomal dominant disease belonging to Tri-Nucleotide repeat
expansion (TNRE) disease
• Expanded CAG repeats coding for polyQ stretch in the htt gene are the main cause of disease.
• Glutamine is a rich metabolite that plays an important role in inflammatory responses.
• Glutamate is a neurotransmitter and the glutamine glutamate shuttle is affected in case of the
Huntington’s disease.
• Glutamine also plays a key role in the activation of the mTOR pathway

MODELS ORGANISMS USED IN HUNTINGTON’S DISEASE RESULTS:

25Q 72Q
STUDY
CTRL (BY4742) CTRL: Yeast BY4742 strain

ΔGCV1

Yeast Cell lines C.elegans Zebra fish Mice Primates

WHY YEAST? ΔGDH1
o Single system
o Easy for genetic manipulations
o Very few variables for studies which makes the study easier. ΔSNZ1
o Helps in minimizing the use of the animal models
Very short life cycle so studying the progeny for disease takes less time
ΔSNO1

OBJECTIVES: ΔTOR1
• How does glutamine modulate the huntingtin protein aggregation?
GCV1: subunit of glycine cleavage complex
• Can glutamine and glutamate-mediated amyloid formation be reduced? CTRL+72Q ΔGLT1 (KO)+72Q
90000
80000
70000
60000

• Showing modulation in the pathway and its effect in the protein aggregation using the knockout 72Q
50000
40000
30000
20000
10000

yeast models and metabolites(inhibitors) 0

250000 1 2

200000

Glutamine(5mM) Florescence in GLT1

150000

100000

compared to CTRL
METHODOLOGY:
50000

0
400000 1 2
350000
300000

Glutamate(5mM)
250000
200000
150000
100000
50000
72Q
0
1 2

Glutamine(5mM)+ 300000
290000
280000

Nicotinic acid
270000
260000
250000
Glutamine
240000

(100uM) ctrl type

yeast

CTRL+72Q ΔGDH1 (KO)+72Q

300000
Glutamate
290000

72Q
280000

Culture
270000
260000

Revival
250000
240000

the Transfor Quantific Glutamine+NA

ctrl type
yeast

of the
400,000

plasmid mation of ation of Glutamate(5mM)

300,000

Midi- wild type 200,000

containin Quantific yeast the 100,000

prep for and Fluoresce 0

1 2

g bacteria ation cells fluoresce

plasmid knockout nce
(25Q and using using the nce
72Q
extractio
Nanodrop
yeast
electropor
imaging
observed CONCLUSION
n strains
obtained ation using • Glutamine is a potential metabolite for increasing the protein aggregation. The knockout yeast
from the
from add method ImageJ
stock models have shown increased mutant huntingtin aggregation in presence of glutamine.
gene
• Glutamate is a critical metabolite in clearing protein aggregation.
• α-ketoglutarate might be a reason for amyloidogenesis.

ACKNOWLEDGMENTS REFERENCES
• Bhagawan Sri Sathya Sai Baba • The global prevalence of Huntington's disease (image courtesy: https://huntingtonstudygroup.org )
• Disease Biology Lab (Dr.S Venketesh and Reasearch scholars) • Ha, A. D., & Fung, V. S. C. (2012). Huntington’s disease. In Current Opinion in Neurology (Vol. 25, Issue 4, pp. 491–498).
• Department of Biosciences https://doi.org/10.1097/WCO.0b013e3283550c97
• Central Research Instruments Facility (CRIF), SSSIHL • Gois, A. M., Mendonça, D. M. F., Freire, M. A. M., & Santos, J. R. (2020). in Vitro and in Vivo Models of Amyotrophic Lateral Sclerosis: an
• Funding agencies Updated Overview. Brain Research Bulletin, 159(November 2019), 32–43. https://doi.org/10.1016/j.brainresbull.2020.03.012
• Sri Sai Sanwid Pradhan • Novak, M. J. U., & Tabrizi, S. J. (2010a). Clinical Review Huntington’s disease. British Medical Journal, 341(July), 34–40
• Sri Shakthi Thangavel SK
• Classmates