Molecular Cloning And Functional Characterization Of Gallus Autophagy Related Gene12 (gaATG12) And Autophagy Related Gene16L1 (gaATG16L1)

Autophagy has been proved to be a conservation mechanism of cellular defense.In mammals and yeasts,it has been reported that ATG12 and ATG16L1 feature significant role in innate immune responses.While the function of ATG12 and ATG16L1 in gallus(gaATG12 and gaATG16L1),a natural host for Newcastle Disease,remains unclear.Previous study confirmed the critical role of autophagy in NDV infection.It is reported that NDV triggered autophagy,resulting in enhanced virus replication.Here we studied the role of gaATG12 and gaATG16L1 in NDV.In this pilot study,full-length of gaATG12 and gaATG16L1 were cloned from chicken kidney.Results showed that gaATG12 was 441 bp in length,and coded a UBQ superfamily structural protein that could help ATG12 combinating with ATG5.While gaATG16L1 was1875 bp in length including a WD40 repeat sequence that worked in the final step of autophagysome.To better understand whether gaATG12 and gaATG16L1 take part in host immune responses,we analysed the expression and variation during NDV infection.We first discovered gaATG12 and gaATG16L1 were expressed across a broad spectrum in chicken by tissue analysis.Additionally,a direct correlation between gaATG12 and gaATG16L1 and cellular inflammatory cytokines expression were found in DF-1 cells.Overexpressed gaATG12 and gaATG16L1 in DF-1 cells showed lower level expression of inflammatory cytokines,including IFN-?,IFN-? and IL-1?,IL-6,IL-8,IL-10,IL-18 induced by NDV comparing with empty vector.Besides,IFN-? was induced by gaATG16L1 in DF-1 cells.Yet IFN-? was susppressed when si RNA-ATG16L1 used before incubating N DV.And most of cytokines tested were upgraded in si RNA experiment.Meanwhile,virus titers were tested with overexpressing gaATG12 and gaATG16L1 or si RNA treatmentsby contrast with empty vector,and results showed that gaATG12 and gaATG16L1 might up-regulate NDV replication.These findings shed new light on the crucial role played by gaATG12 and gaATG16L1 in gallus innate immune system during NDV infection.And leads us to consider moreabout the impact ofhost to the pathogenic process of virus.In conclusion,we confirmed that gaATG12 and gaATG16L1 wereimportant for N DV infection.More importantly,for the first time,we used the chicken model to stud y ATG12 and ATG16L1.O ur studies provided insights into virus-host interaction and the development of antiviral strategies against avain virus infection.