Characterization of the molecular basis of group II intron RNA recognition by CRS1-CRM domains.
CRM (chloroplast RNA splicing and ribosome maturation) is a recently recognized RNA-binding domain of ancient origin that has been retained in eukaryotic genomes only within the plant lineage. Whereas in bacteria CRM domains exist as single domain proteins involved in ribosome maturation, in plants they are found in a family of proteins that contain between one and four repeats. Several members of this family with multiple CRM domains have been shown to be required for the splicing of specific plastidic group II introns. Detailed biochemical analysis of one of these factors in maize, CRS1, demonstrated its high affinity and specific binding to the single group II intron whose splicing it facilitates, the plastid-encoded atpF intron RNA. Through its association with two intronic regions, CRS1 guides the folding of atpF intron RNA into its predicted "catalytically active" form. To understand how multiple CRM domains cooperate to achieve high affinity sequence-specific binding to RNA, we analyzed the RNA binding affinity and specificity associated with each individual CRM domain in CRS1; whereas CRM3 bound tightly to the RNA, CRM1 associated specifically with a unique region found within atpF intron domain I. CRM2, which demonstrated only low binding affinity, also seems to form specific interactions with regions localized to domains I, III, and IV. We further show that CRM domains share structural similarities and RNA binding characteristics with the well known RNA recognition motif domain.
Keren
Ido
I
Institute of Plant Sciences, Agricultural Research Organization, Bet Dagan 50250, Israel.
Klipcan
Liron
L
Bezawork-Geleta
Ayenachew
A
Kolton
Max
M
Shaya
Felix
F
Ostersetzer-Biran
Oren
O
D017398:Alternative Splicing; D005982:Glutathione Transferase / Q000378:metabolism; D017665:Hydroxyl Radical; D007438:Introns; D007700:Kinetics; D009687:Nuclear Proteins / Q000378:metabolism*; D009690:Nucleic Acid Conformation; D010940:Plant Proteins / Q000378:metabolism*; D018087:Plastids / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D012313:RNA / Q000737:chemistry*; D012326:RNA Splicing; D000072260:RNA Splicing Factors; D016601:RNA-Binding Proteins / Q000378:metabolism*; D012270:Ribosomes / Q000737:chemistry*; D003313:Zea mays / Q000378:metabolism*
C436683:CRS1 protein, Zea mays; D009687:Nuclear Proteins; D010940:Plant Proteins; D000072260:RNA Splicing Factors; D016601:RNA-Binding Proteins; D017665:Hydroxyl Radical; D012313:RNA; D005982:Glutathione Transferase