Barthet Michelle M, Pierpont Christopher L, Tavernier Emilie-Katherine
Department of Biology Coastal Carolina University Conway SC USA.
Division of Biological Sciences University of Montana Missoula MT USA.
Plant Direct. 2020 Mar 13;4(3):e00208. doi: 10.1002/pld3.208. eCollection 2020 Mar.
Maturases are prokaryotic enzymes that aid self-excision of introns in precursor RNAs and have evolutionary ties to the nuclear spliceosome. Both the mitochondria and chloroplast, due to their prokaryotic origin, encode a single intron maturase, MatR for the mitochondria and MatK for the chloroplast. MatK is proposed to aid excision of seven different chloroplast group IIA introns that reside within precursor RNAs for essential elements of chloroplast function. We have developed an in vitro activity assay to test chloroplast group IIA intron excision. Using this assay, we demonstrate self-excision of the group IIA intron of the second intron of and the group IIA intron of . We further show that the addition of heterologously expressed MatK protein increases efficiency of group IIA intron self-splicing for the second intron of but not the group IIA intron of . These data, to our knowledge, provide the first direct evidence of MatK's maturase activity.
成熟酶是原核生物酶,可协助前体RNA中内含子的自我切除,并且与核剪接体存在进化联系。线粒体和叶绿体由于其原核生物起源,各自编码一种内含子成熟酶,线粒体的是MatR,叶绿体的是MatK。MatK被认为有助于切除七种不同的叶绿体IIA组内含子,这些内含子存在于叶绿体功能必需元件的前体RNA中。我们开发了一种体外活性测定法来测试叶绿体IIA组内含子的切除。使用该测定法,我们证明了[具体基因名称]第二个内含子的IIA组内含子和[具体基因名称]的IIA组内含子的自我切除。我们进一步表明,添加异源表达的MatK蛋白可提高[具体基因名称]第二个内含子的IIA组内含子自我剪接效率,但不能提高[具体基因名称]的IIA组内含子的效率。据我们所知,这些数据提供了MatK成熟酶活性的首个直接证据。