1-Title: Sequence analysis and molecular characterization of TLR4 gene of camel (Camelus dromedarius)
Authors: Jyoti Choudhary, Mahender Milind, Mukul Purva, Aarti Nirwan, Monika Shekhawat and BN Shringi
Source: Ruminant Science (2022)-11(2):253-260.
How to cite this manuscript: Choudhary Jyoti, Milind Mahender, Purva Mukul, Nirwan Aarti, Shekhawat Monika and Shringi BN (2022). Sequence analysis and molecular characterization of TLR4 gene of camel (Camelus dromedarius). Ruminant Science 11(2):253-260.
Toll-like receptor 4 (TLR4) is essential for initiating the innate immune response to lipopolysaccharide from Gram-negative bacteria by acting as a signal-transducing receptor. At present, there is a paucity of information about most of the dromedary camel (Camelus dromedarius) TLRs. We isolated the complementary DNA (cDNA) by RT-PCR and analysed the TLR4 sequence characters by bioinformatics tool. It was located on the 4th chromosome which had 3 exons. The complete mRNA sequence of the gene has been submitted in NCBI under accession number (MN807289), which was made up of 2936 bases long contig with 2526 long open reading frame (ORF) translated into 841 amino acids (aa) and consist of 20 types of aa. The camel TLR4 sequence had 97.03 per cent identity with the predicted TLR4 protein of dromedary camel with twenty-five additional AA which was justified by the newly updated predicted sequence (XM-010998309.2). TLR4 gene showed the highest degree of aa identity with pigs (83.59 %) whereas, lowest with the rabbit (69.64 %). The protein domain analysis predicted by bioinformatics consists of a transmembrane helices domain, eighteen leucine-rich repeat (LRR) domains, and a Toll/Interleukin-1 receptor (TIR) domain. The TIR domain for downstream signalling transduction was most conservative than other domains after the alignment of protein with different species.
Arbour NC, Lorenz E, Schutte BC, Zabner J, Kline JN, Jones M, Frees K, Watt JL and Schwartz DA (2000). TLR4 mutations are associated with endotoxin hyporesponsiveness in humans. Nature Genetics 25(2):187-191.
Baker, D and Sali, A (2001). Protein structure prediction and structural genomics. Science 294 (5540):93-96.
Branger J, Knapp S, Weijer S, Leemans JC, Pater JM, Speelman P, Florquin S and van der Poll T (2004). Role of toll-like receptor 4 in Gram-positive and Gram-negative pneumonia in mice. Infection and Immunity 72(2):788-94.
Campos MA, Rosinha GM, Almeida IC, Salgueiro XS, Jarvis BW, Splitter GA, Qureshi N, Bruna-Romero O, Gazzinelli RT and Oliveira SC (2004). Role of toll-like receptor 4 in induction of cell-mediated immunity and resistance to Brucella abortus infection in mice. Infection and Immunity 72(1):176-86.
Chang JS, Russell GC, Jann O, Glass EJ, Werling D and Haig DM (2009). Molecular cloning and characterization of Toll-like receptors 1-10 in sheep. Veterinary Immunology and Immunopathology 127(1-2):94-105.
Dahiya SS, Nagarajan G, Bharti VK, Swami SK, Mehta SC, Tuteja FC, Narnaware SD and Patil N (2014). Sequence analysis of the Toll-like receptor 2 gene of old-world camels. Journal of Advanced Research 5(6):695-704.
Janssens S and Beyaert R (2003). Role of Toll-like receptors in pathogen recognition. Clinical Microbiology Reviews 16(4):637-646.
Kang JY and Lee JO (2011). Structural biology of the toll-like receptor family. Annual Review of Biochemistry 80:917-941.
Kanzler H, Barrat FJ, Hessel EM and Coffman RL (2007). Therapeutic targeting of innate immunity with Toll-like receptor agonists and antagonists. Nature Medicine 13(5):552-559.
Kiechl S, Lorenz E, Reindl M, Wiedermann CJ, Oberhollenzer F, Bonora E, Willeit J and Schwartz DA (2002). Toll-like receptor 4 polymorphisms and atherogenesis. New England Journal of Medicine 347(3):185-192.
Lorenz E, Mira JP, Frees KL and Schwartz DA (2002). Relevance of mutations in the TLR4 receptor in patients with gram-negative septic shock. Archives of Internal Medicine 162(9):1028-1032.
Manolakis AC, Kapsoritakis AN, Tiaka EK, Sidiropoulos A, Gerovassili A, Satra M, Vamvakopoulou D, Tsiopoulos F, Papanas N, Skoularigis I and Potamianos SP (2011). TLR4 gene polymorphisms: Evidence for protection against type 2 diabetes but not for diabetes-associated ischaemic heart disease. European Journal of Endocrinology 165(2):261-267.
O’Neill LA and Bowie AG (2007). The family of five: TIR-domain-containing adaptors in Toll-like receptor signalling. Nature Reviews Immunology 7(5):353-364.
Pohar J, Pirher N, Bencina M, Mancek-Keber M and Jerala, R (2014). The ectodomain of TLR3 receptor is required for its plasma membrane translocation. PloS One 9(3):e92391.
Radstake TR, Franke B, Hanssen S, Netea MG, Welsing P, Barrera P, Joosten LA, van Riel PL and van den Berg WB (2004). The Toll like receptor 4 Asp299Gly functional variant is associated with decreased rheumatoid arthritis disease susceptibility but does not influence disease severity and/or outcome. Arthritis and Rheumatism 50(3):999-1001.
Raja A, Vignesh AR, Mary BA, Tirumurugaan KG, Raj GD, Kataria R, Mishra BP and Kumanan K (2011). Sequence analysis of Toll-like receptor genes 1-10 of goat (Capra hircus). Veterinary Immunology and Immunopathology 140(3-4):252-258.
Smith PD, Shimamura M, Musgrove LC, Dennis EA, Bimczok D, Novak L, and Smythies LE (2014). Cytomegalo virus enhances macrophage TLR expression and MyD88-mediated signal transduction to potentiate inducible inflammatory responses. Journal of Immunology 193(11):5604-5612.
Song M, Jin J, Lim JE, Kou J, Pattanayak A, Rehman JA, Kim HD, Tahara K, Lalonde R and Fukuchi KI (2011). TLR4 mutation reduces microglial activation, increases Aâ deposits and exacerbates cognitive deficits in a mouse model of Alzheimer’s disease. Journal of Nneuroinflammation 8(1):1-14
Suthar D, Purva M, Gahlot K, Deora A and Maherchandani S (2019). Sequence analysis of toll-like receptor 1 gene of camel (Camelus dromedarius). Journal of Camel Practice and Research 26(1):41-47.
Tulic MK, Hurrelbrink RJ, Prele CM, Laing IA, Upham JW, Le Souef P, Sly PD and Holt PG (2007). TLR4 polymorphisms mediate impaired responses to respiratory syncytial virus and lipopolysaccharide. Journal of Immunology 179(1):132-40.
Vinkler M, Bainova H and Bryja J (2014). Protein evolution of Toll-like receptors 4, 5 and 7 within Galloanserae birds. Genetics Selection Evolution 46(1):1-12.
Wang X, Xu S, Gao X, Ren H and Chen J (2007). Genetic polymorphism of TLR4 gene and correlation with mastitis in cattle. Journal of Genetics and Genomics 34(5):406-412.