Kitashiba Hiroyasu, Hao Yu-Jin, Honda Chikako, Moriguchi Takaya
National Institute of Fruit Tree Science, Tsukuba, Ibaraki 305-8605, Japan.
Gene. 2005 Nov 21;361:101-11. doi: 10.1016/j.gene.2005.07.013. Epub 2005 Sep 22.
Three cDNAs with high homology to spermine (Spm) synthases in Arabidopsis were isolated from apple [Malus sylvestris (L.) Mill. var. domestica (Borkh.) Mansf.]. MdACL5-1 and MdACL5-2 have high homology with ACL5 and MdSPMS has high homology with AtSPMS. The similarity of MdSPMS to spermidine synthases (SPDSs) was higher than that of MdACL5s, despite the fact that both are putative Spm synthases. However, MdSPMS could be discriminated from SPDSs by the presence of several characteristic amino acids, i.e., Val-149, Ser-161, Ala-205, and Val-235, in the decarboxylated S-adenosylmethionine (dcSAM)-binding motif of MdSPMS. Both MdACL5-1 and MdSPMS complemented Spm biosynthesis in a yeast mutant deficient in Spm synthase, and ectopic expression of MdACL5-1 in the Arabidopsis dwarf mutant acl5 allowed recovery of the normal phenotype. RNA gel blot analysis showed that MdACL5 and MdSPMS are differentially expressed in tissues and suspension cells. These results suggest that functional MdACL5 and MdSPMS are independently involved in apple fruit development and cell growth.
从苹果[苹果(Malus sylvestris (L.) Mill. var. domestica (Borkh.) Mansf.)]中分离出三个与拟南芥精胺(Spm)合酶具有高度同源性的cDNA。MdACL5-1和MdACL5-2与ACL5具有高度同源性,而MdSPMS与AtSPMS具有高度同源性。尽管MdSPMS和MdACL5s都是推定的Spm合酶,但MdSPMS与亚精胺合酶(SPDSs)的相似性高于MdACL5s。然而,通过MdSPMS的脱羧S-腺苷甲硫氨酸(dcSAM)结合基序中存在几个特征性氨基酸,即Val-149、Ser-161、Ala-205和Val-235,可以将MdSPMS与SPDSs区分开来。MdACL5-1和MdSPMS都在缺乏Spm合酶的酵母突变体中补充了Spm生物合成,并且MdACL5-1在拟南芥矮化突变体acl5中的异位表达使正常表型得以恢复。RNA凝胶印迹分析表明,MdACL5和MdSPMS在组织和悬浮细胞中差异表达。这些结果表明,功能性的MdACL5和MdSPMS独立参与苹果果实发育和细胞生长。