Molecular cloning and characterization of autophagy-related gene TmATG8 in Listeria-invaded hemocytes of Tenebrio molitor.
Macroautophagy (hereinafter called autophagy) is a highly regulated process used by eukaryotic cells to digest portions of the cytoplasm that remodels and recycles nutrients and disposes of unwanted cytoplasmic constituents. Currently 36 autophagy-related genes (ATG) and their homologs have been characterized in yeast and higher eukaryotes, including insects. In the present study, we identified and functionally characterized the immune function of an ATG8 homolog in a coleopteran insect, Tenebrio molitor (TmATG8). The cDNA of TmATG8 comprises of an ORF of 363 bp that encodes a protein of 120 amino acid residues. TmATG8 transcripts are detected in all the developmental stages analyzed. TmAtg8 protein contains a highly conserved C-terminal glycine residue (Gly116) and shows high amino acid sequence identity (98%) to its Tribolium castaneum homolog, TcAtg8. Loss of function of TmATG8 by RNAi led to a significant increase in the mortality rates of T. molitor larvae against Listeria monocytogenes. Unlike dsEGFP-treated control larvae, TmATG8-silenced larvae failed to turn-on autophagy in hemocytes after injection with L. monocytogenes. These data suggest that TmATG8 play a role in mediating autophagy-based clearance of Listeria in T. molitor.
Division of Plant Biotechnology, Institute of Environmentally-Friendly Agriculture (IEFA), College of Agriculture and Life Sciences, Chonnam National University, Gwangju, Republic of Korea; Department of Soil Science, Faculty of Agriculture, Sokoine University of Agriculture, P.O. Box 3008, Chuo Kikuu, Morogoro, Tanzania.
Hamisi
H
Tindwa
Division of Plant Biotechnology, Institute of Environmentally-Friendly Agriculture (IEFA), College of Agriculture and Life Sciences, Chonnam National University, Gwangju, Republic of Korea.
Yong Hun
YH
Jo
Division of Plant Biotechnology, Institute of Environmentally-Friendly Agriculture (IEFA), College of Agriculture and Life Sciences, Chonnam National University, Gwangju, Republic of Korea; Trident School of Biotech Sciences, Trident Academy of Creative Technology, Chandrasekharpur, Bhubaneswar 751024, Odisha, India.
Bharat Bhusan
BB
Patnaik
Department of Life Science and Biotechnology, College of Natural Sciences, Soonchunhyang University, Asan City, Republic of Korea.
Yong Seok
YS
Lee
Department of Biology Education, Chungbuk National University, 410 Seongbong Road, Seowon-gu, Cheongju, Chungbuk 361-763, Republic of Korea.
Sang Sun
SS
Kang
Division of Plant Biotechnology, Institute of Environmentally-Friendly Agriculture (IEFA), College of Agriculture and Life Sciences, Chonnam National University, Gwangju, Republic of Korea. Electronic address: hanys@jnu.ac.kr.
Yeon Soo
YS
Han
D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D001343:Autophagy / Q000235:genetics; D000071190:Autophagy-Related Protein 8 Family; D058491:Bacterial Load / Q000235:genetics; D003001:Cloning, Molecular; D006434:Hemocytes / Q000382:microbiology / Q000502:physiology*; D008089:Listeria monocytogenes / Q000276:immunology*; D008088:Listeriosis / Q000276:immunology*; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D034741:RNA, Small Interfering / Q000235:genetics; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013712:Tenebrio / Q000276:immunology*
D048868:Adaptor Proteins, Signal Transducing; D000071190:Autophagy-Related Protein 8 Family; C498698:GABARAPL2 protein, human; D008840:Microfilament Proteins; D034741:RNA, Small Interfering
ATG8; Autophagy; Innate immunity; Listeria monocytogenes; RNA interference; Tenebrio molitor