The carbonic anhydrase isoforms of Chlamydomonas reinhardtii: Intracellular location, expression, and physiological roles

James V. Moroney, Louisiana State University
Yunbing Ma, Louisiana State University
Wesley D. Frey, Louisiana State University
Katelyn A. Fusilier, Louisiana State University
Trang T. Pham, Louisiana State University
Tiffany A. Simms, Louisiana State University
Robert J. DiMario, Louisiana State University
Jing Yang, Louisiana State University
Bratati Mukherjee, Louisiana State University

Abstract

Aquatic photosynthetic organisms, such as the green alga Chlamydomonas reinhardtii, respond to low CO2 conditions by inducing a CO 2 concentrating mechanism (CCM). Carbonic anhydrases (CAs) are important components of the CCM. CAs are zinc-containing metalloenzymes that catalyze the reversible interconversion of CO2 and HCO 3-. In C. reinhardtii, there are at least 12 genes that encode CA isoforms, including three alpha, six beta, and three gamma or gamma-like CAs. The expression of the three alpha and six beta genes has been measured from cells grown on elevated CO2 (having no active CCM) versus cells growing on low levels of CO2 (with an active CCM) using northern blots, differential hybridization to DNA chips and quantitative RT-PCR. Recent RNA-seq profiles add to our knowledge of the expression of all of the CA genes. In addition, protein content for some of the CA isoforms was estimated using antibodies corresponding to the specific CA isoforms: CAH1/2, CAH3, CAH4/5, CAH6, and CAH7. The intracellular location of each of the CA isoforms was elucidated using immunolocalization and cell fractionation techniques. Combining these results with previous studies using CA mutant strains, we will discuss possible physiological roles of the CA isoforms concentrating on how these CAs might contribute to the acquisition and retention of CO2 in C. reinhardtii. © Springer Science+Business Media B.V. 2011.