Addepalli Balasubrahmanyam, Meeks Lisa R, Forbes Kevin P, Hunt Arthur G
Plant Physiology/Biochemistry/Molecular Biology Program and Department of Agronomy, University of Kentucky, 301A Plant Science Building, 1405 Veterans Road, Lexington, KY 40546-0312, USA.
Biochim Biophys Acta. 2004 Aug 12;1679(2):117-28. doi: 10.1016/j.bbaexp.2004.06.001.
The Arabidopsis thaliana genome possesses four genes whose predicted products are similar to eukaryotic poly(A) polymerases from yeasts and animals. These genes are all expressed, as indicated by RT/PCR and Northern blot analysis. The four Arabidopsis PAPs share a conserved N-terminal catalytic core with other eukaryotic enzymes, but differ substantially in their C-termini. Moreover, one of the four Arabidopsis enzymes is significantly shorter than the other three, and is more divergent even within the conserved core of the protein. Nonetheless, the protein encoded by this gene, when produced in and purified from E. coli, possesses nonspecific poly(A) polymerase activity. Genes encoding these Arabidopsis PAPs give rise to a number of alternatively spliced mRNAs. While the specific nature of the alternative splicing varied amongst these three genes, mRNAs from the three "larger" genes could be alternatively spliced in the vicinity of the 5th and 6th introns of each gene. Interestingly, the patterns of alternative splicing vary in different tissues. The ubiquity of alternative splicing in this gene family, as well as the differences in specific mechanisms of alternative processing in the different genes, suggests an important function for alternatively spliced PAP mRNAs in Arabidopsis.
拟南芥基因组中有四个基因,其预测产物与来自酵母和动物的真核多聚腺苷酸聚合酶相似。如逆转录/聚合酶链反应(RT/PCR)和Northern印迹分析所示,这些基因均有表达。四种拟南芥多聚腺苷酸聚合酶(PAP)与其他真核酶一样,在N端有一个保守的催化核心,但C端差异很大。此外,四种拟南芥酶中的一种明显短于其他三种,甚至在蛋白质的保守核心区域内差异也更大。尽管如此,该基因编码的蛋白质在大肠杆菌中表达并纯化后,具有非特异性多聚腺苷酸聚合酶活性。编码这些拟南芥PAP的基因会产生许多可变剪接的mRNA。虽然这三个基因的可变剪接具体性质各不相同,但来自三个“较大”基因的mRNA可在每个基因的第5和第6个内含子附近进行可变剪接。有趣的是,可变剪接模式在不同组织中有所不同。该基因家族中可变剪接的普遍性,以及不同基因可变加工具体机制的差异,表明可变剪接的PAP mRNA在拟南芥中具有重要功能。