Identification and characterization of multidomain monothiol glutaredoxin 3 from diploblastic Hydra

Area of Science:

• Molecular Biology
• Cell Biology
• Biochemistry

Background:

• Glutaredoxins (Grxs) are intracellular antioxidants controlling cell proliferation and survival.
• Grxs are classified into dithiol (Class I, CXXC active site) and monothiol (Class II, single cysteine active site) isoforms.
• Hydra vulgaris possesses multiple Grx isoforms, including the previously characterized dithiol HvGrx1.

Purpose of the Study:

• To clone, express, and characterize a novel multidomain monothiol glutaredoxin-3 (HvGrx3) from Hydra vulgaris.
• To investigate the functional role of HvGrx3 in cellular processes like proliferation, migration, and oxidative stress response.

Main Methods:

• Molecular cloning and in-silico analysis of HvGrx3.
• Expression analysis in Hydra vulgaris whole body and H2O2-treated samples.
• Transfection of HvGrx3 into HCT116 human colon cancer cells for functional studies.

Main Results:

• HvGrx3 is a multidomain monothiol Grx, distinct from HvGrx1, with two Grx domains and an N-terminal thioredoxin-like domain.
• HvGrx3 expression is widespread in Hydra and significantly upregulated by hydrogen peroxide.
• Transfected HCT116 cells expressing HvGrx3 exhibited enhanced proliferation, migration, and tolerance to oxidative stress.

Conclusions:

• HvGrx3 is a functional monothiol glutaredoxin involved in cellular proliferation, migration, and oxidative stress management.
• HvGrx3 represents a distinct Grx isoform with potentially significant roles in cellular homeostasis and cancer biology.