Knock-out of a putative transporter results in altered blue-light signalling in Chlamydomonas.

Abstract

Nitrogen starvation and blue light are the two environmental cues that control sexual differentiation in Chlamydomonas reinhardtii. Insertional mutagenesis was applied to generate mutants that still require nitrogen starvation as the initiating signal for gametogenesis but were no longer dependent on irradiation. In one mutant analysed, sequences adjacent to the site of insertion were cloned and used for the isolation of a genomic clone that, upon transformation, could complement the mutant phenotype. The gene identified (LRG6) encodes two mRNAs that appear to be the products of differential splicing. The two putative gene products derived from these mRNAs differ in their C-terminal ends. Both predicted gene products exhibit multiple hydrophobic domains with alpha-helical secondary structure typical for integral membrane proteins. These proteins may form pores, and may function as transporters of as-yet unknown substrates. Since rendering the LRG6 gene non-functional resulted in light-independence of gamete formation, it is suggested that this transporter may inhibit signal flux from the photoreceptor to target genes - either directly by its activity or indirectly by serving as a scaffold for signalling proteins. Shutting off this transporter may be required for the activation of signal flux in this pathway. This concept is supported by the observed reduction in LRG6 mRNA levels during the first phase of gametic differentiation.