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The unicellular green algae  Chlamydomonas reinhardtii  has long been studied for its unique fermentation pathways and has been evaluated as a candidate organism for biofuel production. Fermentation in  C .  reinhardtii  is facilitated by a network of three predominant pathways producing four major byproducts: formate, ethanol, acetate and hydrogen. Previous microarray studies identified many genes as being highly up-regulated during anaerobiosis. For example,  hybrid cluster protein 4  ( HCP4 ) was found to be one of the most highly up-regulated genes under anoxic conditions. Hybrid cluster proteins have long been studied for their unique spectroscopic properties, yet their biological functions remain largely unclear. To probe its role during anaerobiosis,  HCP4  was silenced using artificial microRNAs ( ami-hcp4 ) followed by extensive phenotypic analyses of cells grown under anoxic conditions. Both the expression of key fermentative enzymes and their respective metabolites were significantly altered in  ami-hcp4 , with nitrogen uptake from the media also being significantly different than wild-type cells. The results strongly suggest a role for  HCP4  in regulating key fermentative and nitrogen utilization pathways.

The Involvement of hybrid cluster protein 4, HCP4, in Anaerobic Metabolism in Chlamydomonas reinhardtii