Regulación de la actividad de la fosforilación oxidativa mitocondrial mediada por modificaciones post-traduccionales

Manchón Campillo, Teresa (2021). Regulación de la actividad de la fosforilación oxidativa mitocondrial mediada por modificaciones post-traduccionales. Proyecto Fin de Carrera / Trabajo Fin de Grado, E.T.S. de Ingeniería Agronómica, Alimentaria y de Biosistemas (UPM), Madrid.

Description

Title: Regulación de la actividad de la fosforilación oxidativa mitocondrial mediada por modificaciones post-traduccionales
Author/s:
  • Manchón Campillo, Teresa
Contributor/s:
  • Cuezva Marcos, José Manuel
  • Torres Lacruz, Miguel Ángel
Item Type: Final Project
Degree: Grado en Biotecnología
Date: June 2021
Subjects:
Faculty: E.T.S. de Ingeniería Agronómica, Alimentaria y de Biosistemas (UPM)
Department: Biotecnología - Biología Vegetal
Creative Commons Licenses: Recognition - No derivative works - Non commercial

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Abstract

The regulation of mitochondrial oxidative phosphorylation (OXPHOS) is crucial in physiological conditions as well as in pathologies such as in cancer. Post-translational modifications of the OXPHOS system remain largely unknown but play a key role in its regulation, especially protein phosphorylation. Protein kinase A (PKA) is one of the enzymes regulating the phosphorylation of OXPHOS proteins, what results in increasing the activity of some complexes of the electron transport chain (complexes I and IV) and in the inactivation of the ATPase Inhibitory Factor 1 (IF1), preventing its binding to the ATP synthase. In order to assist PKA in the phosphorylation of IF1, A-kinase anchoring proteins (AKAPs) have been postulated as accessory proteins of the process. Furthermore, the activity of PKA is dependent on cyclic AMP (cAMP), synthesized within the mitochondria by the Soluble Adenylyl Cyclase (sAC). Herein, we studied the impact of the AKAPs AKAP1, WAVE1 and SKIP on the phosphorylation of IF1 in colon and breast cancer cells. Moreover, we studied the potential implication of sAC in the phosphorylation of different OXPHOS complexes. To these aims, first we carried out the silencing of the AKAPs in both cell lines and studied the phosphorylation status of IF1 by two- dimensional electrophoresis. Next, we developed a sAC knockout cell line, using CRISPR-Cas9 technology, and studied the serine phosphorylation status of Complexes I, IV and V after their immunocapture. We observed that silencing of AKAP1 and WAVE1 partially prevents the phosphorylation of IF1 in breast cancer cells, whereas it has no impact in colon cancer cells. However, the absence of IF1 phosphorylation does not affect basal mitochondrial respiration. Furthermore, we observed that sAC ablated cells do not modify the phosphorylation status of complexes I and IV in response to treatment with the membrane permeable db-cAMP - activator of PKA- and nebivolol, a β1-adrenergic blocker an inhibitor of PKA signaling. In contrast, these treatments respectively promoted or prevented the phosphorylation of one subunit of complex V. Altogether, these results highlight the tissue-specific relevance of protein phosphorylation to control the activity of OXPHOS and stress the need of future studies to investigate the role of accessory proteins to understand the role of PKA within the mitochondria.

More information

Item ID: 69181
DC Identifier: https://oa.upm.es/69181/
OAI Identifier: oai:oa.upm.es:69181
Deposited by: Biblioteca ETSI Agrónomos
Deposited on: 02 Dec 2021 14:15
Last Modified: 02 Feb 2022 23:30
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