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Identification, expression and subcellular localization of Orc1 in the microsporidian Nosema bombycis
Fuzhen Sun1, Guanyu Zhu1, Ping He1
1Jiangsu University of Science and Technology, Zhenjiang 212018, Jiangsu Province, China.
Abstract:
As a typical species of microsporidium, Nosema bombycis is the pathogen causing the pébrine disease of silkworm. Rapid proliferation of N. bombycis in host cells requires replication of genetic material. As eukaryotic origin recognition protein, origin recognition complex (ORC) plays an important role in regulating DNA replication, and Orc1 is a key subunit of the origin recognition complex. In this study, we identified the Orc1 in the microsporidian N. bombycis (NbOrc1) for the first time. The NbOrc1 gene contains a complete ORF of 987 bp in length that encodes a 328 amino acid polypeptide. Indirect immunofluorescence results showed that NbOrc1 were colocalized with Nbactin and NbSAS-6 in the nuclei of N. bombycis. Subsequently, we further identified the interaction between the NbOrc1 and Nbactin by CO-IP and Western blot. These results imply that Orc1 may be involved in the proliferation of the microsporidian N. bombycis through interacting with actin.
Insights
Researchers identified the origin recognition complex subunit 1 (Orc1) in Nosema bombycis, a silkworm pathogen. This protein, NbOrc1, interacts with actin, suggesting a role in parasite proliferation and DNA replication.
Area of Science:
- Microbiology and Parasitology
- Molecular Biology
- Silkworm Pathology
Background:
- Nosema bombycis is a microsporidian parasite causing pébrine disease in silkworms.
- Parasite proliferation necessitates DNA replication, a process regulated by the origin recognition complex (ORC) in eukaryotes.
- Orc1 is a crucial subunit of the ORC, involved in initiating DNA replication.
Purpose of the Study:
- To identify and characterize the Orc1 homolog in Nosema bombycis (NbOrc1).
- To investigate the cellular localization and potential interactions of NbOrc1.
- To explore the role of NbOrc1 in the proliferation of N. bombycis.
Main Methods:
- Gene identification and sequencing of NbOrc1.
- Indirect immunofluorescence microscopy to determine NbOrc1 localization.
- Co-immunoprecipitation (CO-IP) and Western blot assays to confirm protein interactions.
Main Results:
- The NbOrc1 gene was identified, encoding a 328-amino acid polypeptide.
- NbOrc1 was found to colocalize with Nbactin and NbSAS-6 within the nucleus of N. bombycis.
- Interaction between NbOrc1 and Nbactin was confirmed through CO-IP and Western blot.
Conclusions:
- NbOrc1 is a functional Orc1 subunit in Nosema bombycis.
- The interaction between NbOrc1 and Nbactin suggests a novel role for Orc1 in microsporidian proliferation.
- NbOrc1 may be a potential target for controlling pébrine disease by interfering with parasite DNA replication and proliferation.
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