Title The EH domain of EHD3 is required, but not sufficient, to cause its aggregation
Title (croatian) EH domena EHD3 proteina je potrebna, ali ne i dovoljna, za njegovu agregaciju
Author Ema Ukić
Mentor Nicholas Bradshaw (mentor)
Committee member Nicholas Bradshaw (predsjednik povjerenstva)
Committee member Miranda Mladinić Pejatović (član povjerenstva)
Committee member Mladen Merćep (član povjerenstva)
Committee member Jelena Ban (član povjerenstva)
Granter University of Rijeka (Faculty of Biotechnology and Drug Development) Rijeka
Defense date and country 2023-09-27, Croatia
Scientific / art field, discipline and subdiscipline BIOTECHNICAL SCIENCES Biotechnology
Abstract Schizophrenia is a serious mental illness characterised by impaired
perception of reality and changes in behaviour. The molecular background of
schizophrenia is not yet fully known. In addition to genetic and
environmental factors, protein aggregation is being researched as one of the
possible causes of schizophrenia. Protein aggregation occurs when, affected
by endogenous and external stresses, cellular mechanisms get disrupted,
resulting in protein misfolding and protein aggregates accumulating. EHdomain containing 3 (EHD3) is a protein that has been found to aggregate in
the brains of schizophrenia patients. EHD3 is a protein involved in endocytic
trafficking and recycling as well as D1 receptor internalization. Full length
EHD3 has been shown to aggregate in SH-SY5Y cells, while two constructs
lacking the C-terminus of the protein, EHD3 amino acids 1-434 and 1-399,
did not aggregate. This implied that amino acids 435-535, representing its
EH domain, were required for aggregation. Also, full length EHD3 has been
shown to induce EHD3 1-399 to aggregation when co-expressed. In this
thesis we aimed to determine if the EH domain aggregates independently
and if it would co-aggregate when co-expressed with full length EHD3. When
expressed individually, EHD3 435-535 did not aggregate, implying that the
EH domain is not sufficient for EHD3 aggregation by itself. When coexpressed with full length EHD3, EHD3 1-399 and EHD3 1-434 the EH
domain also did not aggregate, which might imply that while the EH domain
is necessary for the initialization of EHD3 aggregation, other structural
regions like the helical and/or the G domain interact and recruit other
molecules that make up the aggregates. Future research is needed to
determine which EHD3 regions interact with the EH domain causing the
protein to aggregate and co-aggregate in the cell.
Abstract (croatian) Šizofrenija je ozbiljna mentalna bolest koju karakteriziraju narušena
percepcija stvarnosti i promjene u ponašanju pojedinca. Molekularni
mehanizmi šizofrenije još uvijek nisu dovoljno istraženi. Uz poznate
genetske i okolišne čimbenike, agregacija proteina se istražuje kao
potencijalni uzrok šizofrenije. Do agregacije proteina dolazi kada se uslijed
različitih endogenih i mehaničkih stresova naruše stanični mehanizmi što
rezultira pogrešnim slaganjem proteina i nakupljanjem njihovih agregata.
EH-domain containing 3 (EHD3) protein jedan je od proteina koji su
identificirani kao agregirajući u mozgovima pacijenata oboljelih od
šizofrenije. To je protein koji sudjeluje u endosomalnom transportu i
recikliranju te u internalizaciji D1 receptora. Dokazano je kako EHD3
agregira u SH-SY5Y staničnoj liniji, dok konstrukti kojima nedostaje Cterminalna regija, EHD3 1-434 i EHD3 1-399 ne agregiraju, Ovo upućuje
kako su aminokiseline 435-535, koje predstavljaju EH domenu proteina,
nužne za njegovu agregaciju. Također je pokazano kako EHD3 može
inducirati agregaciju EHD3 1-399 kada se zajedno eksprimiraju u stanici. U
ovom radu nastojali smo pokazati agregira li EH domena samostalno u
stanici, te hoće li ko-agregirati s cijelim EHD3 proteinom ukoliko se zajedno
eksprimiraju. EHD3 435-535 (sadrži isključivo EH domenu) ne agregira kada
se samostalno eksprimira u stanici, niti su cijeli EHD3, EHD3 1-399 i EHD3
1-434 inducirali njegovu agregaciju. Ovo upućuje da je EH domena ključna
za inicijalizaciju agregacije EHD3 proteina, no druge strukturalne regije
proteina, poput zavojnica i G domene, su potrebne za interakciju i
regrutaciju drugih molekula koje izgrađuju agregat. Daljnja istraživanja
potrebna su kako bi odredili koje to regije stupaju u interakciju s EH
domenom rezultirajući agregacijom i ko-agregacijom EHD3 proteina.
Keywords
schizophrenia
protein aggregation
EHD3
Keywords (croatian)
šizofrenija
agregacija proteina
EHD3
Language english
URN:NBN urn:nbn:hr:193:134090
Study programme Title: Biotechnology in medicine Study programme type: university Study level: graduate Academic / professional title: magistar/magistra biotehnologije u medicini (magistar/magistra biotehnologije u medicini)
Type of resource Text
File origin Born digital
Access conditions Open access
Terms of use
Created on 2023-10-11 10:47:57