
NSF Org: |
MCB Division of Molecular and Cellular Biosciences |
Recipient: |
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Initial Amendment Date: | July 31, 2018 |
Latest Amendment Date: | July 13, 2021 |
Award Number: | 1817338 |
Award Instrument: | Continuing Grant |
Program Manager: |
Wilson Francisco
wfrancis@nsf.gov (703)292-7856 MCB Division of Molecular and Cellular Biosciences BIO Directorate for Biological Sciences |
Start Date: | August 1, 2018 |
End Date: | July 31, 2023 (Estimated) |
Total Intended Award Amount: | $899,995.00 |
Total Awarded Amount to Date: | $974,931.00 |
Funds Obligated to Date: |
FY 2020 = $359,998.00 FY 2021 = $74,936.00 |
History of Investigator: |
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Recipient Sponsored Research Office: |
55 LAKE AVE N WORCESTER MA US 01655-0002 (508)856-2119 |
Sponsor Congressional District: |
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Primary Place of Performance: |
55 North Lake Ave Worcester MA US 01655-0002 |
Primary Place of
Performance Congressional District: |
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Unique Entity Identifier (UEI): |
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Parent UEI: |
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NSF Program(s): |
Molecular Biophysics, Cross-BIO Activities |
Primary Program Source: |
01002021DB NSF RESEARCH & RELATED ACTIVIT 01002122DB NSF RESEARCH & RELATED ACTIVIT |
Program Reference Code(s): |
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Program Element Code(s): |
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Award Agency Code: | 4900 |
Fund Agency Code: | 4900 |
Assistance Listing Number(s): | 47.074 |
ABSTRACT
Multimeric ATPases drive the mechanics of biology, facilitating critical biosynthetic pathways and controlling cellular and viral dynamics. An exceptionally powerful multimeric ATPase is the terminase motor that pumps DNA into viral capsids during assembly of dsDNA viruses. These extremely powerful terminase enzymes provide unique opportunities for uncovering how the force of motor output is governed. Understanding of motor mechanism could lead to development of new nanomaterials that respond to the environment, or nanodevices for targeted delivery of nucleic acids. Furthermore, these motors are of interest in development of novel sequencing technologies using nanopores for reading out nucleic acid sequence. This innovative research and outreach platform will explore the terminase's nanotechnological potential, while also providing educational opportunities at the high school and graduate level.
The structural mechanism of terminase motors at the atomic level will be elucidated using a combination of x-ray crystallography, cryo-electron microscopy and molecular modelling to determine three-dimensional structures of terminases and their complexes. New structures will be used to generate hypotheses for motor mechanism, which will then be tested using innovative biochemical and biophysical assays. These studies will complement the investigators research on AAA+ ATPases to reveal how ATPases couple chemical energy to mechanical motion, providing broad and novel insights into the mechanisms, regulation, and evolution of molecular motors.
This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.
PUBLICATIONS PRODUCED AS A RESULT OF THIS RESEARCH
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PROJECT OUTCOMES REPORT
Disclaimer
This Project Outcomes Report for the General Public is displayed verbatim as submitted by the Principal Investigator (PI) for this award. Any opinions, findings, and conclusions or recommendations expressed in this Report are those of the PI and do not necessarily reflect the views of the National Science Foundation; NSF has not approved or endorsed its content.
We are interested in how biological materials can survive harsh environments. Biotechnologies have revolutionized many aspects of our lives, from medicine to manufacturing, but we have only begun to realize their full potential. Most products available only work in a narrow range of temperatures and conditions. For example, vaccines and other therapeutics usually need to be refrigerated until they are administered, which costs energy and greatly limits where we can ship them and where patients can be treated. In addition, many industrial processes require high temperatures or harsh chemicals, which would destroy the biotechnologies we might use to improve their efficiency. Many organisms have evolved to thrive in these exact conditions, and studying how their molecules work can show us general principles for designing more durable biotechnologies.
We chose to study a virus found in hot springs and has adapted to thrive at extremely hot temperatures. This virus is a “bacteriophage,” which means that it infects bacteria. Bacteriophages are common in every environment on earth, including our own bodies, and much of our understanding of biology has been learned from studying them.
We discovered how its shell, or capsid, can withstand high temperature. Viruses use these protein shells to protect their DNA, and if the shell is broken the virus cannot survive. We used incredibly powerful microscopes to look at the individual atoms that make up the capsid. By comparing our data to viruses that are not able to survive high temperatures, we were able to identify characteristics that make this capsid heat resistant, as well as ways that the shell can encapsulate larger cargo. This opens the door to use the capsid as a building block for new nanotechnologies. For instance, protein engineers may choose to attach molecules to the outside of the capsid to create new vaccines or load capsids with different genetic material to create new therapies.
We also studied how the virus gets its DNA inside of the capsid. First, we learned how the virus recognizes its own DNA rather than all the other DNA that is abundant inside of a cell. We went on to show how the virus moves DNA from outside the capsid to inside the capsid. While humans do not move our DNA inside of capsids, our cells constantly need to recognize DNA and move DNA and other molecules around, and our findings help us understand how many of these processes may work.
In addition to a protein shell, this virus also has a long tail that it uses to infect its prey. Many other bacteriophages have similar tails, but this virus has the longest tail ever observed. Like the capsid, the tail is extremely heat resistant. We used the same microscopes to view the molecules of the tails, allowing us to determine how it withstands high temperatures. While the capsid is shaped like a soccer ball, the tail is shaped like a long pipe. This means that they can be adapted for different nanotechnologies. While the capsid is rigid and a good shape for carrying cargo inside or for organizing molecules arranged like a ball, the tail is flexible and a good shape for making wires or organizing molecules like beads on a string or in a woven mesh.
Like all viruses, this bacteriophage needs to get its DNA into its prey in order to infect it. Most bacteriophages simply inject their DNA into their prey without protecting it from the cell’s immune system. Using the same high-magnification microscopes, we discovered that this bacteriophage creates a bubble around its DNA by hijacking components of the cell’s outer covering. This kind of bubble has never been observed in bacteria before, and we think that it protects the bacteriophage DNA from the prey’s immune system, which has evolved to detect and destroy virus DNA.
This groundbreaking research was completed by students within our university, as well as those we have hosted from other schools and from our local community. A student from Puerto Rico whose education was disrupted by Hurricane Maria and the COVID pandemic and a local high school student carried out important experiments to determine how the virus infects its prey. In addition to making discoveries, these students learned research skills and advanced their own careers.
It is equally important to share our science and passion for science with our local community. Dr. Kelch advises students from Wachusett Regional High School on Tuesday nights (October through March), offering encouragement and guidance on their science fair projects. Many of Dr. Kelch’s advisees placed in the Massachusetts State Science Fair, including the 2023 Grand Prize winner. We participated in the Skype-a-Scientist program which matches scientists with middle and high school classrooms across the country, as well as the ScienceLive program at UMMS which provides structured outreach to local middle school students.
Last Modified: 11/29/2023
Modified by: Brian Kelch
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