Tatematsu Kiyoshi, Kumagai Satoshi, Muto Hideki, Sato Atsuko, Watahiki Masaaki K, Harper Reneé M, Liscum Emmanuel, Yamamoto Kotaro T
Division of Biological Sciences, Graduate School of Environmental Earth Science, Hokkaido University, Sapporo 060-0810, Japan.
Plant Cell. 2004 Feb;16(2):379-93. doi: 10.1105/tpc.018630. Epub 2004 Jan 16.
We have isolated a dominant, auxin-insensitive mutant of Arabidopsis thaliana, massugu2 (msg2), that displays neither hypocotyl gravitropism nor phototropism, fails to maintain an apical hook as an etiolated seedling, and is defective in lateral root formation. Yet other aspects of growth and development of msg2 plants are almost normal. These characteristics of msg2 are similar to those of another auxin-insensitive mutant, non-phototropic hypocotyl4 (nph4), which is a loss-of-function mutant of AUXIN RESPONSE FACTOR7 (ARF7) (Harper et al., 2000). Map-based cloning of the MSG2 locus reveals that all four mutant alleles result in amino acid substitutions in the conserved domain II of an Auxin/Indole-3-Acetic Acid protein, IAA19. Interestingly, auxin inducibility of MSG2/IAA19 gene expression is reduced by 65% in nph4/arf7. Moreover, MSG2/IAA19 protein binds to the C-terminal domain of NPH4/ARF7 in a Saccharomyces cerevisiae (yeast) two-hybrid assay and to the whole latter protein in vitro by pull-down assay. These results suggest that MSG2/IAA19 and NPH4/ARF7 may constitute a negative feedback loop to regulate differential growth responses of hypocotyls and lateral root formation.
我们分离出了拟南芥的一个显性生长素不敏感突变体——massugu2(msg2),该突变体既不表现下胚轴向重力性也不表现向光性,在黄化幼苗期无法维持顶端弯钩,并且侧根形成存在缺陷。然而,msg2植株生长和发育的其他方面几乎正常。msg2的这些特征与另一个生长素不敏感突变体——非向光性下胚轴4(nph4)相似,nph4是生长素响应因子7(ARF7)的功能缺失突变体(Harper等人,2000年)。对MSG2基因座进行图位克隆发现,所有四个突变等位基因都导致生长素/吲哚-3-乙酸蛋白IAA19保守结构域II中的氨基酸替换。有趣的是,在nph4/arf7中,MSG2/IAA19基因表达的生长素诱导性降低了65%。此外,在酿酒酵母双杂交试验中,MSG2/IAA19蛋白与NPH4/ARF7的C末端结构域结合,在体外通过下拉试验与后者的整个蛋白结合。这些结果表明,MSG2/IAA19和NPH4/ARF7可能构成一个负反馈环,以调节下胚轴的差异生长反应和侧根形成。