Genomic organization, sequence characterization and expression analysis of Tenebrio molitor apolipophorin-III in response to an intracellular pathogen, Listeria monocytogenes

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• 作者：Ju Young Noh ; Bharat Bhusan Patnaik ; Hamisi Tindwa ; Gi Won Seo ; Dong Hyun Kim ; Hongray Howrelia Patnaik ; Yong Hun Jo ; Yong Seok Lee ; Bok Luel Lee ; Nam Jung Kim ; Yeon Soo Han
• 关键词：LTA ; Lipoteichoic acid ; LPS ; Lipopolyssacharide ; PAMP ; Pathogen associated molecular patterns ; PPO ; Prophenoloxidase ; ATCC ; American Type Culture Collection ; BHI ; Brain&ndash ; Heart infusion ; TB ; Terrific broth ; ENA-EBI ; European Nucleotide Archive-European Bioinformatics Institute ; SMART ; Simple modular architecture research tool ; NCBI ; National Centre for Biotechnology Information ; NJ ; Neighbor-joining ; ML ; Maximum likelihood ; NMR ; Nuclear magnetic resonance ; qRTPCR ; Quantitative real-time PCR
• 刊名：Gene
• 出版年：25 January, 2014
• 年：2014
• 卷：534
• 期：2
• 页码：204-217
• 全文大小：2542 K

文摘

Apolipophorin III (apoLp-III) is a well-known hemolymph protein having a functional role in lipid transport and immune response of insects. We cloned full-length cDNA encoding putative apoLp-III from larvae of the coleopteran beetle, Tenebrio molitor (TmapoLp-III), by identification of clones corresponding to the partial sequence of TmapoLp-III, subsequently followed with full length sequencing by a clone-by-clone primer walking method. The complete cDNA consists of 890 nucleotides, including an ORF encoding 196 amino acid residues. Excluding a putative signal peptide of the first 20 amino acid residues, the 176-residue mature apoLp-III has a calculated molecular mass of 19,146 Da. Genomic sequence analysis with respect to its cDNA showed that TmapoLp-III was organized into four exons interrupted by three introns. Several immune-related transcription factor binding sites were discovered in the putative 5鈥?flanking region. BLAST and phylogenetic analyses reveal that TmapoLp-III has high sequence identity (88%) with Tribolium castaneum apoLp-III but shares little sequence homologies (< 26%) with other apoLp-IIIs. Homology modeling of Tm apoLp-III shows a bundle of five amphipathic alpha helices, including a short helix 3鈥? The 鈥榟elix-short helix-helix鈥?motif was predicted to be implicated in lipid binding interactions, through reversible conformational changes and accommodating the hydrophobic residues to the exterior for stability. Highest level of TmapoLp-III mRNA was detected at late pupal stages, albeit it is expressed in the larval and adult stages at lower levels. The tissue specific expression of the transcripts showed significantly higher numbers in larval fat body and adult integument. In addition, TmapoLp-III mRNA was found to be highly upregulated in late stages of L. monocytogenes or E. coli challenge. These results indicate that TmapoLp-III may play an important role in innate immune responses against bacterial pathogens in T. molitor.