A nuclear mutation that affects the 3' processing of several mRNAs in Chlamydomonas chloroplasts

Abstract

We previously created and analyzed a Chlamydomonas reinhardtii strain, β26, in which an inverted repeat in the 3' untranslated region of the chloroplast atpB gene was deleted. In this strain, atpB transcripts are unstable and heterogeneous in size, and growth is poor under conditions in which photosynthesis is required. Spontaneous suppressor mutations that allow rapid photosynthetic growth have been identified. One strain, β26S, retains the atpB deletion yet accumulates a discrete and stable atpB transcript as a consequence of a recessive nuclear mutation. Unlike previously isolated Chlamydomonas nuclear mutations that affect chloroplast mRNA accumulation, the mutation in β26S affects several chloroplast transcripts. For example, in the atpA gene cluster, the relative abundance of several messages was altered in a manner consistent with inefficient mRNA 3' end processing. Furthermore, β26S cells accumulated novel transcripts with 3' tarmini in the petD-tmR intergenic region. These transcripts are potential intermediates in 3' end processing. In contrast, no alterations were detected for petD, atpA, or atpB mRNA 5' ends; neither were there gross alterations detected for several other mRNAs, including the wild-type atpB transcript. We suggest that the gene identified by the suppressor mutation encodes a product involved in the processing of monocistronic and polycistronic messages.