Pregled bibliografske jedinice broj: 1208233
Determination of the domain structure of DISC1 in mammalian cells identifies a region crucial for its aggregation in schizophrenia
Determination of the domain structure of DISC1 in mammalian cells identifies a region crucial for its aggregation in schizophrenia // FEBS Open Bio, 12 (Suppl S1)
Lisabon, Portugal, 2022. str. 54-54 (poster, međunarodna recenzija, sažetak, znanstveni)
CROSBI ID: 1208233 Za ispravke kontaktirajte CROSBI podršku putem web obrasca
Naslov
Determination of the domain structure of DISC1 in
mammalian cells identifies a region crucial for
its aggregation in schizophrenia
Autori
Zaharija, Beti ; Sanchez-Pulido, Luis ; Bradshaw, Nicholas J.
Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni
Izvornik
FEBS Open Bio, 12 (Suppl S1)
/ - , 2022, 54-54
Skup
The Biochemistry Global Summit
Mjesto i datum
Lisabon, Portugal, 06.07.2022. - 14.07.2022
Vrsta sudjelovanja
Poster
Vrsta recenzije
Međunarodna recenzija
Ključne riječi
DISC1 ; mental illness ; schizophrenia ; protein aggregation ; domain structure
Sažetak
While the pathology of schizophrenia remains unclear, disrupted protein homeostasis has recently been suggested as its biological basis. Several proteins have now been shown to form insoluble aggregates in this condition, including Disrupted in Schizophrenia 1 (DISC1), a multi- functional scaffolding protein important for neurodevelopment. However, delineation of DISC1 domain structure has only recently been attempted using high-throughput analysis in E. coli, which identified four distinct domains: D, I, S and C. This data was used to confirm and refine the structure of DISC1 in mammalian cells, as well as to identify the region of DISC1 responsible for its aggregation. To do so, DISC1 domain borders were refined by combining previously published bioinformatics data with the recent empirical data. Constructs encoding variants of three domains were cloned, with borders modified based on theoretical predictions. These were transfected into HEK293 cells and tested for stability via a proteasome inhibition assay. Modified versions of the D and C domains showed improved stability over their empirical counterparts, while the I region may not represent a stable folded domain by itself. The D, S and C domains were further shown to be functional in isolation via interactions with known binding partners. To investigate which structural region might be responsible for its aggregation, DISC1 fragments were expressed in SH- SY5Y cells, with localization patterns viewed by fluorescent microscopy. Single domains showed diffuse cytoplasmic localization, while the combination of domains D and I showed clear aggregation. We therefore hypothesized that the unstructured region between domains D and I is responsible for DISC1 aggregation propensity, which was verified using further truncation constructs. Together, our data will aid in generation of high-quality animal models to study the mechanism and behavioral consequences of DISC1 aggregation, and its relevance for schizophrenia.
Izvorni jezik
Engleski
Znanstvena područja
Biotehnologija u biomedicini (prirodno područje, biomedicina i zdravstvo, biotehničko područje)
POVEZANOST RADA
Projekti:
--IP-2018-01-9424 - Istraživanje shizofrenije kroz ekspresiju netopivih proteina (CandidIskren) (Bradshaw, Nicholas James) ( CroRIS)
--DOK-2018-09-5395 - Istraživanje shizofrenije kroz ekspresiju netopivih proteina (CandidIskren) (Bradshaw, Nicholas James) ( CroRIS)
Ustanove:
Sveučilište u Rijeci - Odjel za biotehnologiju