Dark-induced mRNA instability involves RNase E/G-type endoribonuclease cleavage at the AU-box and SD sequences in cyanobacteria |
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Authors: | Yoshinao Horie Yoko Ito Miyuki Ono Naoko Moriwaki Hideki Kato Yuriko Hamakubo Tomoki Amano Masaaki Wachi Makoto Shirai Munehiko Asayama |
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Institution: | (1) Laboratory of Molecular Genetics, School of Agriculture, Ibaraki University, Ami, Inashiki, Ibaraki 300-0393, Japan;(2) Department of Bioengineering, Tokyo Institute Technology, 4259 Nagatsuta, Midori-ku, Yokohama 226-8503, Japan |
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Abstract: | Light-responsive gene expression is crucial to photosynthesizing organisms. Here, we studied functions of cis-elements (AU-box and SD sequences) and a trans-acting factor (ribonuclease, RNase) in light-responsive expression in cyanobacteria. The results indicated that AU-rich nucleotides
with an AU-box, UAAAUAAA, just upstream from an SD confer instability on the mRNA under darkness. An RNase E/G homologue,
Slr1129, of the cyanobacterium Synechocystis sp. strain PCC 6803 was purified and confirmed capable of endoribonucleolytic cleavage at the AU- (or AG)-rich sequences
in vitro. The cleavage depends on the primary target sequence and secondary structure of the mRNA. Complementation tests using
Escherichia coli rne/rng mutants showed that Slr1129 fulfilled the functions of both the RNase E and RNase G. An analysis of systematic mutations
in the AU-box and SD sequences showed that the cis-elements also affect significantly mRNA stability in light-responsive genes. These results strongly suggested that dark-induced
mRNA instability involves RNase E/G-type cleavage at the AU-box and SD sequences in cyanobacteria. The mechanical impact and
a possible common mechanism with RNases for light-responsive gene expression are discussed.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. |
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Keywords: | Cyanobacteria Endoribonuclease Degradosome Light-responsive gene psbA 5′ -UTR |
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