PPR-SMR protein SOT1 has RNA endonuclease activity.
Numerous attempts have been made to identify and engineer sequence-specific RNA endonucleases, as these would allow for efficient RNA manipulation. However, no natural RNA endonuclease that recognizes RNA in a sequence-specific manner has been described to date. Here, we report that SUPPRESSOR OF THYLAKOID FORMATION 1 (SOT1), anArabidopsispentatricopeptide repeat (PPR) protein with a small MutS-related (SMR) domain, has RNA endonuclease activity. We show that the SMR moiety of SOT1 performs the endonucleolytic maturation of 23S and 4.5S rRNA through the PPR domain, specifically recognizing a 13-nucleotide RNA sequence in the 5' end of the chloroplast 23S-4.5S rRNA precursor. In addition, we successfully engineered the SOT1 protein with altered PPR motifs to recognize and cleave a predicted RNA substrate. Our findings point to SOT1 as an exciting tool for RNA manipulation.
Photosynthesis Research Center, Key Laboratory of Photobiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China.|College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China.
Wen
W
Zhou
Photosynthesis Research Center, Key Laboratory of Photobiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China.
Qingtao
Q
Lu
Photosynthesis Research Center, Key Laboratory of Photobiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China.|College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China.
Qingwei
Q
Li
Photosynthesis Research Center, Key Laboratory of Photobiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China.|College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China.
Lei
L
Wang
Photosynthesis Research Center, Key Laboratory of Photobiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China.
Shunhua
S
Ding
Photosynthesis Research Center, Key Laboratory of Photobiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China.
Aihong
A
Zhang
Photosynthesis Research Center, Key Laboratory of Photobiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China.
Xiaogang
X
Wen
Photosynthesis Research Center, Key Laboratory of Photobiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China.
Lixin
L
Zhang
Photosynthesis Research Center, Key Laboratory of Photobiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China; lucm@ibcas.ac.cn.|National Center for Plant Gene Research, Beijing 100093, China.
Congming
C
Lu
D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D002736:Chloroplasts / Q000235:genetics / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D004722:Endoribonucleases / Q000235:genetics / Q000378:metabolism*; D005818:Genetic Engineering; D008565:Membrane Proteins / Q000378:metabolism; D014176:Protein Biosynthesis; D018747:RNA, Chloroplast / Q000378:metabolism*; D012338:RNA, Ribosomal, 23S / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D020524:Thylakoids / Q000378:metabolism*
D029681:Arabidopsis Proteins; D008565:Membrane Proteins; D018747:RNA, Chloroplast; D012338:RNA, Ribosomal, 23S; D011994:Recombinant Proteins; C512069:THF1 protein, Arabidopsis; C538752:pentatricopeptide repeat protein, Arabidopsis; D004722:Endoribonucleases
PPR-SMR protein; RNA endonuclease; photosynthesis; rRNA biogenesis