Henry Isabelle M, Wilkinson Mark D, Hernandez J Marcela, Schwarz-Sommer Zsuzsanna, Grotewold Erich, Mandoli Dina F
Department of Biology, University of Washington, Box 355325, 1521 NE Pacific Street, Seattle, WA 98195-5325, USA.
BMC Plant Biol. 2004 Mar 12;4:3. doi: 10.1186/1471-2229-4-3.
Acetabularia acetabulum is a giant unicellular green alga whose size and complex life cycle make it an attractive model for understanding morphogenesis and subcellular compartmentalization. The life cycle of this marine unicell is composed of several developmental phases. Juvenile and adult phases are temporally sequential but physiologically and morphologically distinct. To identify genes specific to juvenile and adult phases, we created two subtracted cDNA libraries, one adult-specific and one juvenile-specific, and analyzed 941 randomly chosen ESTs from them.
Clustering analysis suggests virtually no overlap between the two libraries. Preliminary expression data also suggests that we were successful at isolating transcripts differentially expressed between the two developmental phases and that many transcripts are specific to one phase or the other. Comparison of our EST sequences against publicly available sequence databases indicates that ESTs from the adult and the juvenile libraries partition into different functional classes. Three conserved sequence elements were common to several of the ESTs and were also found within the genomic sequence of the carbonic anhydrase1 gene from A. acetabulum. To date, these conserved elements are specific to A. acetabulum.
Our data provide strong evidence that adult and juvenile phases in A. acetabulum vary significantly in gene expression. We discuss their possible roles in cell growth and morphogenesis as well as in phase change. We also discuss the potential role of the conserved elements found within the EST sequences in post-transcriptional regulation, particularly mRNA localization and/or stability.
伞藻是一种巨大的单细胞绿藻,其大小和复杂的生命周期使其成为理解形态发生和亚细胞区室化的有吸引力的模型。这种海洋单细胞的生命周期由几个发育阶段组成。幼年和成年阶段在时间上是连续的,但在生理和形态上是不同的。为了鉴定幼年和成年阶段特有的基因,我们创建了两个消减cDNA文库,一个是成年特异性的,一个是幼年特异性的,并分析了从中随机选择的941个EST。
聚类分析表明两个文库之间几乎没有重叠。初步表达数据还表明,我们成功地分离了两个发育阶段差异表达的转录本,并且许多转录本是一个阶段或另一个阶段特有的。将我们的EST序列与公开可用的序列数据库进行比较表明,来自成年和幼年文库的EST分为不同的功能类别。几个EST共有三个保守序列元件,并且在伞藻碳酸酐酶1基因的基因组序列中也发现了这些元件。迄今为止,这些保守元件是伞藻特有的。
我们的数据提供了有力的证据,表明伞藻的成年和幼年阶段在基因表达上有显著差异。我们讨论了它们在细胞生长和形态发生以及相变中的可能作用。我们还讨论了在EST序列中发现的保守元件在转录后调控,特别是mRNA定位和/或稳定性中的潜在作用。