Weis V M, Reynolds W S
Department of Zoology, Oregon State University, Corvallis, OR, 97331 USA.
Physiol Biochem Zool. 1999 May-Jun;72(3):307-16. doi: 10.1086/316674.
Endosymbiotic dinoflagellates resident within cnidarian hosts are extremely productive primary producers. This high productivity may be due in part to an inorganic carbon transport system, present in host tissue, that accelerates carbon delivery to the algae. The enzyme carbonic anhydrase (CA; EC 4.2.1.1) has been shown to be important in this transport system in a variety of tropical symbiotic cnidarians. This study extends the examination of CA to a temperate anemone, Anthopleura elegantissima, and documents symbiosis-enhanced production of CA at the biochemical and molecular level. Depending on light availability, A. elegantissima can occur naturally with (symbiotic) or without (aposymbiotic) dinoflagellates, making it an ideal study organism for symbiosis-enhanced gene expression. We compared (1) CA activities, (2) quantities of CA using an anti-human CA immunoprobe, and (3) quantities of transcript using a semiquantitative PCR in symbiotic versus aposymbiotic A. elegantissima host tissue. Amounts of activity, enzyme, and transcript were greatly enhanced in symbiotic animals compared with aposymbiotic animals. This is the first direct evidence that the presence of symbionts affects the expression of a host cnidarian gene. In addition, we report a full-length A. elegantissima CA cDNA sequence, obtained from subcloned reverse transcriptase-PCR products, and its relatedness to alpha-CAs from a variety of other metazoa, including higher vertebrates.
生活在刺胞动物宿主内的内共生甲藻是极具生产力的初级生产者。这种高生产力可能部分归因于宿主组织中存在的一种无机碳转运系统,该系统加速了碳向藻类的输送。碳酸酐酶(CA;EC 4.2.1.1)已被证明在多种热带共生刺胞动物的这种转运系统中起着重要作用。本研究将对CA的研究扩展到一种温带海葵——优美列指海葵,并在生化和分子水平上记录了共生增强的CA产生情况。根据光照条件,优美列指海葵可以自然地与(共生)或不与(非共生)甲藻共生,这使其成为研究共生增强基因表达的理想生物。我们比较了共生与非共生的优美列指海葵宿主组织中的(1)CA活性、(2)使用抗人CA免疫探针检测的CA量,以及(3)使用半定量PCR检测的转录本量。与非共生动物相比,共生动物中的活性、酶和转录本量大大增加。这是共生体的存在影响宿主刺胞动物基因表达的首个直接证据。此外,我们报告了从亚克隆的逆转录酶 - PCR产物中获得的优美列指海葵CA全长cDNA序列,以及它与包括高等脊椎动物在内的多种其他后生动物的α - CA的亲缘关系。