Poynter Sarah J, Monjo Andrea L, Micheli Gabriella, DeWitte-Orr Stephanie J
Department of Biology, University of Waterloo, 200 University Ave W, Waterloo, ON N2L 3G1, Canada.
Department of Biology, Wilfrid Laurier University, 75 University Ave W, Waterloo, ON N2L 3C5, Canada.
Dev Comp Immunol. 2017 Dec;77:95-105. doi: 10.1016/j.dci.2017.07.021. Epub 2017 Jul 22.
Class A scavenger receptors (SR-As) are a family of surface-expressed receptors who bind a wide range of polyanionic ligands including bacterial components and nucleic acids and play a role in innate immunity. Macrophage receptor with collagenous structure (MARCO) is a SR-A family member that has been studied in mammals largely for its role in binding bacteria. To date there is little information about SR-As in general and MARCO specifically in fish, particularly what ligands individual SR-A family members bind remains largely unknown. In the present study two novel rainbow trout MARCO transcript variants have been identified and their sequence and putative protein domains have been analyzed. When overexpressed in CHSE-214, a cell line that appears to lack functional scavenger receptors, GFP-tagged rtMARCO-1 and rtMARCO-2 were able to bind gram-positive, and gram-negative bacteria of both mammalian and aquatic sources. rtMARCO appears to bind bacteria via its scavenger receptor cysteine-rich (SRCR) domain, because SRCR deleted rtMARCO-1 and -2 were unable to bind bacteria. rtMARCO did not show any binding to the yeast cell wall component zymosan or to double-stranded (ds)RNA. This is the first time rainbow trout MARCO sequences have been identified and the first in-depth study exploring their ligand binding profile. This study provides novel insight into the role of rainbow trout MARCO in bacterial innate immunity.
A类清道夫受体(SR-As)是一类表面表达的受体家族,它们能结合多种多阴离子配体,包括细菌成分和核酸,并在先天免疫中发挥作用。具有胶原结构的巨噬细胞受体(MARCO)是SR-A家族的成员,在哺乳动物中,主要因其在结合细菌方面的作用而得到研究。迄今为止,关于鱼类中一般的SR-As,特别是MARCO的信息很少,尤其是单个SR-A家族成员结合何种配体在很大程度上仍然未知。在本研究中,已鉴定出两种新的虹鳟MARCO转录变体,并对其序列和推测的蛋白质结构域进行了分析。当在似乎缺乏功能性清道夫受体的CHSE-214细胞系中过表达时,绿色荧光蛋白标记的rtMARCO-1和rtMARCO-2能够结合革兰氏阳性菌以及来自哺乳动物和水生来源的革兰氏阴性菌。rtMARCO似乎通过其富含半胱氨酸的清道夫受体(SRCR)结构域结合细菌,因为缺失SRCR的rtMARCO-1和-2无法结合细菌。rtMARCO对酵母细胞壁成分酵母聚糖或双链(ds)RNA没有任何结合作用。这是首次鉴定出虹鳟MARCO序列,也是首次对其配体结合谱进行深入研究。本研究为虹鳟MARCO在细菌先天免疫中的作用提供了新的见解。