Cristin-resultat-ID: 2045728
Sist endret: 24. august 2022, 14:34
Resultat
Vitenskapelig foredrag
2022

Identification of interaction partners of the Albino3 insertases Alb3a and Alb3b in the diatom Phaeodactylum tricornutum

Bidragsytere:
  • Manuel Serif
  • Per Winge og
  • Marianne Nymark

Presentasjon

Navn på arrangementet: NBS Contact Meeting 2022
Sted: Tromsø
Dato fra: 10. februar 2022
Dato til: 13. februar 2022

Arrangør:

Arrangørnavn: NBS

Om resultatet

Vitenskapelig foredrag
Publiseringsår: 2022

Beskrivelse Beskrivelse

Tittel

Identification of interaction partners of the Albino3 insertases Alb3a and Alb3b in the diatom Phaeodactylum tricornutum

Sammendrag

Diatoms are a group of eukaryotic microalgae of high ecological importance, contributing to up to 20% of oxygen production worldwide. They are of commercial interest for potential biotechnological applica- tions, e.g. for use as fish feed in aquaculture due to their lipid profile or for bio-remediation of wastewa- ter. Pigment-binding proteins of the light harvesting antenna are inserted into the thylakoid membrane via the chloroplast signal recognition particle pathway. Previous studies in plants and green algae have shown that knockout of genes of this pathway can result in strains with a truncated light harvesting antenna. These strains are valuable for biotechnological approaches as they may grow better in dense bio-reactor cultures. Two isoforms of the Albino3 insertase Alb3, named Alb3a and Alb3b, had been identified in the model diatom Phaeodactylum tricornutum. Knockout of Alb3a was attempted, but no bi -allelic loss-of-function mutant could be obtained, indicating that Alb3a is an essential gene. In contrast, knockout of Alb3b resulted in a pronounced change of coloration of the cells from brown to green and a strong decrease in the main light-harvesting antenna proteins was observed. However, growth rates were found to be reduced. Thus, further genes involved in this pathway need to be identified. There- fore, we are employing a proximity labeling approach to identify interaction partners of the two Alb3 proteins. Phaeodactylum tricornutum strains expressing fusion proteins of the respective Alb3 isoform and the peroxidase APEX2 were generated. These strains were then acclimated to medium light condi- tions, followed by a shift to low light conditions to induce Alb3 activity. Upon addition of the membrane- permeable substrate biotin-phenol, APEX2 biotinylates proteins in close proximity, which are then affini- ty purified and identified by mass spectrometry. Potential interaction partners identified will then be confirmed and characterized by generation of knockout strains.

Bidragsytere

Manuel Philipp Serif

Bidragsyterens navn vises på dette resultatet som Manuel Serif
  • Tilknyttet:
    Forfatter
    ved Institutt for biologi ved Norges teknisk-naturvitenskapelige universitet

Per Winge

  • Tilknyttet:
    Forfatter
    ved Institutt for biologi ved Norges teknisk-naturvitenskapelige universitet

Marianne Nymark

  • Tilknyttet:
    Forfatter
    ved Institutt for biologi ved Norges teknisk-naturvitenskapelige universitet
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