PUBLICATION
Elucidation of the mechanism of nuclear localization of Mexican tetra Neu4 via bipartite nuclear localization signal and less conserved regions
- Authors
- Hyodo, T., Honda, A., Yamate, S., Kubo, Y., Komatsu, M., Shiozaki, K.
- ID
- ZDB-PUB-230425-55
- Date
- 2023
- Source
- Biochimie 212: 123-134 (Journal)
- Registered Authors
- Shiozaki, Kazuhiro
- Keywords
- none
- MeSH Terms
-
- Amino Acid Sequence
- Animals
- Cell Nucleus/metabolism
- Characidae*
- Neuraminidase/chemistry
- Nuclear Localization Signals*/genetics
- PubMed
- 37094779 Full text @ Biochimie
Citation
Hyodo, T., Honda, A., Yamate, S., Kubo, Y., Komatsu, M., Shiozaki, K. (2023) Elucidation of the mechanism of nuclear localization of Mexican tetra Neu4 via bipartite nuclear localization signal and less conserved regions. Biochimie. 212:123-134.
Abstract
Nuclear sialoglycans are minor components in the nucleus, and their biological significance was not well understood. Recently, Nile tilapia Neu4 sialidase (OnNeu4) was identified and reported as the first nuclear sialidase in vertebrates. Although OnNeu4 possesses the nuclear localization signal (NLS) required for nuclear localization, other fish Neu4 sialidases, such as zebrafish and medaka, also possess NLS, but their subcellular localization is not nucleus. To understand the nuclear localization mechanism of fish Neu4, we focused on Mexican tetra Neu4 (AmNeu4), which, unlike Neu4 in other fishes, has a bipartite NLS. AmNeu4 exhibits a wide range of optimal pH and substrate specificity, and its gene expression was specifically detected in the liver, spleen, and gut in adult fish. AmNeu4, like OnNeu4, exhibited nuclear localization, which was attenuated by importin inhibitor, and deletion of the bipartite NLS completely reduced the nuclear localization. In addition, the conjugation of the bipartite NLS of AmNeu4 made GFP show nuclear localization. To understand the mechanism of nuclear localization of AmNeu4 and OnNeu4, we compared fish Neu4 amino acid sequences and focused on the less conserved region of Neu4 sialidase (LCR). LCR-deletion mutants of AmNeu4 and OnNeu4 showed significantly reduced the nuclear localization. The LCR region in AmNeu4 and OnNeu4 possessed consecutive Ser/Thr. The Neu4 mutants in which consecutive Ser/Thr in LCR were changed to Ala or deleted significantly suppressed the nuclear localization. These results suggest that the nuclear localization of Neu4 in tilapia and Mexican tetra may be regulated by NLS and LCR.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping