RAB24 interaction between GOLGA3, USO1 and emerin
Tapio, Johannes (2024-11-12)
RAB24 interaction between GOLGA3, USO1 and emerin
(12.11.2024)
Julkaisu on tekijänoikeussäännösten alainen. Teosta voi lukea ja tulostaa henkilökohtaista käyttöä varten. Käyttö kaupallisiin tarkoituksiin on kielletty.
avoin
Julkaisun pysyvä osoite on:
https://urn.fi/URN:NBN:fi-fe2024112997720
https://urn.fi/URN:NBN:fi-fe2024112997720
Tiivistelmä
RAB proteins are small GTPases responsible for controlling membrane trafficking in eukaryotic
cells. RAB24 is a unique RAB protein, due to its low GTPase activity. It is implicated to function
at later stages of basal autophagy and in the endocytic degradation pathway. It is also required for
normal cell division and plays a role in liver cell homeostasis. A point mutation in RAB24 causes
hereditary ataxia in dogs, potentially through disruptions in the autophagic process. In addition,
increased expression of RAB24 promotes a malignant phenotype of hepatocellular carcinoma
(HCC) cells.
The exact functional mechanisms of RAB24 are still unknown. Critical in this regard is to identify
the proteins that RAB24 interacts with in the cell. Using proximity labeling, the Eskelinen group
discovered new putative RAB24 interaction partners, of which three are studied in this thesis;
Golgin subfamily A member 3 (GOLGA3), General vesicular transport factor p115 (USO1, also
called VDP) and emerin.
The aim of this thesis was to investigate whether GOLGA3, USO1 and emerin physically interact
with RAB24, and whether they co-localize with RAB24 within the cell. We used co
immunoprecipitation (co-IP) to study the physical interaction between the proteins, and
immunofluorescence microscopy to visualize and quantify protein co-localization. Experiments
were conducted using the human liver cancer-derived cell lines Hep G2 and Huh-7. Interaction
between RAB24 and the three candidate proteins could not be confirmed, as the co-IP results
were inconsistent. USO1 was found to partially co-localize with RAB24.
cells. RAB24 is a unique RAB protein, due to its low GTPase activity. It is implicated to function
at later stages of basal autophagy and in the endocytic degradation pathway. It is also required for
normal cell division and plays a role in liver cell homeostasis. A point mutation in RAB24 causes
hereditary ataxia in dogs, potentially through disruptions in the autophagic process. In addition,
increased expression of RAB24 promotes a malignant phenotype of hepatocellular carcinoma
(HCC) cells.
The exact functional mechanisms of RAB24 are still unknown. Critical in this regard is to identify
the proteins that RAB24 interacts with in the cell. Using proximity labeling, the Eskelinen group
discovered new putative RAB24 interaction partners, of which three are studied in this thesis;
Golgin subfamily A member 3 (GOLGA3), General vesicular transport factor p115 (USO1, also
called VDP) and emerin.
The aim of this thesis was to investigate whether GOLGA3, USO1 and emerin physically interact
with RAB24, and whether they co-localize with RAB24 within the cell. We used co
immunoprecipitation (co-IP) to study the physical interaction between the proteins, and
immunofluorescence microscopy to visualize and quantify protein co-localization. Experiments
were conducted using the human liver cancer-derived cell lines Hep G2 and Huh-7. Interaction
between RAB24 and the three candidate proteins could not be confirmed, as the co-IP results
were inconsistent. USO1 was found to partially co-localize with RAB24.