jf7b03734_si_001.pdf (1001.54 kB)
Destruxin A Induces and Binds HSPs in Bombyx mori Bm12 Cells
journal contribution
posted on 2017-10-19, 00:00 authored by Huanhuan Zhang, Weina Hu, Miaomiao Xiao, Shiyi Ou, Qiongbo HuDestruxin
A (DA) is a cyclodepsipeptidic mycotoxin isolated from
the entomopathogenic fungus, Metarhizium anisopliae. It has insecticidal activity against host insect’s innate
immunity system, but the molecular mechanism is not yet elucidated.
In our previous experiment, four HSPs (heat shock proteins, BmHSP70–3,
BmHSP75, BmHSP83, and BmHSCP) were characterized from the specific
protein electrophoretic bands of Bombyx mori Bm12
cell line treated with DA in the test of drug affinity responsive
target stability (DARTS), which implied that these HSPs might be kinds
of DA-affinity proteins, or DA induces them up-regulated expression.
Therefore, in current research, the interactions of DA and HSPs were
explored through analysis of bio-layer interferometry (BLI) employing
FortBio OcteteQK. The expression levels of HSPs genes were surveyed
by quantitative real-time polymerase chain reaction (qPCR). The results
indicated that DA had no interactions with BmHSP70–3, BmHSP75,
and BmHSP83, but had affinity to BmHSCP with a KD value of 88.1 μM, in BLI analysis. However, the expression
levels of all HSPs genes were significantly up-regulated after the
Bm12 cells were treated by DA. In conclusion, DA can induce the four
HSPs expression in Bm12 cells, but DA only binds to BmHSCP. Our research
provides new insights on understanding of the action mechanisms of
destruxins.