Cassidy, Kevin C.
(2021)
Investigating the dynamics of LARP1 DM15: A stalwart of ribosome biogenesis regulation.
Doctoral Dissertation, University of Pittsburgh.
(Unpublished)
Abstract
Eukaryotic cells regulate the synthesis of proteins based on environmental conditions such as nutrient abundance or stress. Ribosome biogenesis is a key node in this regulation. La-related protein 1 (LARP1) has recently been identified as a downstream target of mTORC1 that regulates the translation of terminal oligopyrimidine (TOP) mRNAs, which encode all ribosomal proteins. LARP1 phosphorylation by mTORC1 modulates the association of LARP1 with TOP mRNA. Previous studies support both a repressive and stimulatory role for LARP1 in TOP mRNA translation, and its exact role is still unknown; LARP1 has also been shown to stabilize TOP mRNAs. Fully characterizing the role of LARP1 in translation regulation is vitally important because it is upregulated in several carcinomas and is an attractive pharmacological target. The best strategy to pharmacologically target LARP1 depends on the knowledge of how it affects TOP translation, and consequently, how best to modulate its role in that regulation.
Here we used molecular dynamics simulations to investigate the dynamics of the LARP1 DM15 region and provide insight into the dynamics that govern its role in translation regulation. We found that the open or closed state of the cap-binding pocket is correlated with the secondary structure of an adjacent stretch of residues. Additional LARP1 DM15 pockets were identified that have putative allosteric relationships with either the cap- or +1- binding pockets. Co-evolution analysis reveals a potential autoregulatory mechanism via an interdomain interaction with two of these pockets. A breast cancer-associated LARP1 mutation was introduced into one of the pockets in silico, and molecular dynamics simulations revealed the mutation alters the dynamics of both the cap- and +1-binding pockets, potentially altering the ability to bind mRNAs. Lastly, a combination of in silico and in vitro techniques were used to identify compounds that putatively bind LARP1 DM15. Future experiments could utilize the compound hits to investigate the role of LARP1 in TOP mRNA translation, and optimize them for therapeutic use.
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Details
| Item Type: |
University of Pittsburgh ETD
|
| Status: |
Unpublished |
| Creators/Authors: |
|
| ETD Committee: |
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| Date: |
8 October 2021 |
| Date Type: |
Publication |
| Defense Date: |
22 June 2021 |
| Approval Date: |
8 October 2021 |
| Submission Date: |
5 July 2021 |
| Access Restriction: |
2 year -- Restrict access to University of Pittsburgh for a period of 2 years. |
| Number of Pages: |
187 |
| Institution: |
University of Pittsburgh |
| Schools and Programs: |
Dietrich School of Arts and Sciences > Biological Sciences |
| Degree: |
PhD - Doctor of Philosophy |
| Thesis Type: |
Doctoral Dissertation |
| Refereed: |
Yes |
| Uncontrolled Keywords: |
LARP1, DM15, Cancer, Molecular Dynamics Simulation, TOP mRNA |
| Date Deposited: |
08 Oct 2021 20:21 |
| Last Modified: |
08 Oct 2023 05:15 |
| URI: |
http://d-scholarship.pitt.edu/id/eprint/41392 |
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