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两种羟脯氨酸半乳糖基转移酶GALT5和GALT2在拟南芥阿拉伯半乳聚糖蛋白糖基化、生长和发育过程中发挥作用。

Two Hydroxyproline Galactosyltransferases, GALT5 and GALT2, Function in Arabinogalactan-Protein Glycosylation, Growth and Development in Arabidopsis.

作者信息

Basu Debarati, Wang Wuda, Ma Siyi, DeBrosse Taylor, Poirier Emily, Emch Kirk, Soukup Eric, Tian Lu, Showalter Allan M

机构信息

Molecular and Cellular Biology Program, Department of Environmental and Plant Biology, Ohio University, Athens, Ohio, United States of America.

出版信息

PLoS One. 2015 May 14;10(5):e0125624. doi: 10.1371/journal.pone.0125624. eCollection 2015.

DOI:10.1371/journal.pone.0125624
PMID:25974423
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4431829/
Abstract

Hydroxyproline-O-galactosyltransferase (GALT) initiates O-glycosylation of arabinogalactan-proteins (AGPs). We previously characterized GALT2 (At4g21060), and now report on functional characterization of GALT5 (At1g74800). GALT5 was identified using heterologous expression in Pichia and an in vitro GALT assay. Product characterization showed GALT5 specifically adds galactose to hydroxyproline in AGP protein backbones. Functions of GALT2 and GALT5 were elucidated by phenotypic analysis of single and double mutant plants. Allelic galt5 and galt2 mutants, and particularly galt2 galt5 double mutants, demonstrated lower GALT activities and reductions in β-Yariv-precipitated AGPs compared to wild type. Mutant plants showed pleiotropic growth and development phenotypes (defects in root hair growth, root elongation, pollen tube growth, flowering time, leaf development, silique length, and inflorescence growth), which were most severe in the double mutants. Conditional mutant phenotypes were also observed, including salt-hypersensitive root growth and root tip swelling as well as reduced inhibition of pollen tube growth and root growth in response to β-Yariv reagent. These mutants also phenocopy mutants for an AGP, SOS5, and two cell wall receptor-like kinases, FEI1 and FEI2, which exist in a genetic signaling pathway. In summary, GALT5 and GALT2 function as redundant GALTs that control AGP O-glycosylation, which is essential for normal growth and development.

摘要

羟脯氨酸 - O - 半乳糖基转移酶（GALT）启动阿拉伯半乳聚糖蛋白（AGP）的O - 糖基化。我们之前对GALT2（At4g21060）进行了表征，现在报告GALT5（At1g74800）的功能表征。通过在毕赤酵母中的异源表达和体外GALT测定鉴定出了GALT5。产物表征表明，GALT5特异性地将半乳糖添加到AGP蛋白主链中的羟脯氨酸上。通过单突变体和双突变体植物的表型分析阐明了GALT2和GALT5的功能。与野生型相比，等位基因galt5和galt2突变体，特别是galt2 galt5双突变体，表现出较低的GALT活性以及β - 崖豆素沉淀的AGP减少。突变体植物表现出多效性的生长和发育表型（根毛生长、根伸长、花粉管生长、开花时间、叶片发育、角果长度和花序生长缺陷），在双突变体中最为严重。还观察到了条件性突变体表型，包括盐敏感的根生长和根尖肿胀，以及对β - 崖豆素试剂响应时花粉管生长和根生长的抑制减弱。这些突变体还模拟了AGP、SOS5以及两个细胞壁类受体激酶FEI1和FEI2的突变体，它们存在于一个遗传信号通路中。总之，GALT5和GALT2作为冗余的GALT发挥作用，控制AGP的O - 糖基化，这对正常生长和发育至关重要。

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