Elucidation of structure-function relationships in Methanocaldococcus jannaschii RNase P, a multi-subunit catalytic ribonucleoprotein
dc.contributor.author | Phan, Hong-Duc | en |
dc.contributor.author | Norris, Andrew S. | en |
dc.contributor.author | Du, Chen | en |
dc.contributor.author | Stachowski, Kye | en |
dc.contributor.author | Khairunisa, Bela H. | en |
dc.contributor.author | Sidharthan, Vaishnavi | en |
dc.contributor.author | Mukhopadhyay, Biswarup | en |
dc.contributor.author | Foster, Mark P. | en |
dc.contributor.author | Wysocki, Vicki H. | en |
dc.contributor.author | Gopalan, Venkat | en |
dc.date.accessioned | 2022-11-01T15:08:51Z | en |
dc.date.available | 2022-11-01T15:08:51Z | en |
dc.date.issued | 2022-08-12 | en |
dc.description.abstract | RNase P is a ribonucleoprotein (RNP) that catalyzes removal of the 5 ' leader from precursor tRNAs in all domains of life. A recent cryo-EM study of Methanocaldococcus jannaschii (Mja) RNase P produced a model at 4.6-angstrom resolution in a dimeric configuration, with each holoenzyme monomer containing one RNase P RNA (RPR) and one copy each of five RNase P proteins (RPPs; POP5, RPP30, RPP21, RPP29, L7Ae). Here, we used native mass spectrometry (MS), mass photometry (MP), and biochemical experiments that (i) validate the oligomeric state of the Mja RNase P holoenzyme in vitro, (ii) find a different stoichiometry for each holoenzyme monomer with up to two copies of L7Ae, and (iii) assess whether both L7Ae copies are necessary for optimal cleavage activity. By mutating all kink-turns in the RPR, we made the discovery that abolishing the canonical L7Ae-RPR interactions was not detrimental for RNase P assembly and function due to the redundancy provided by protein-protein interactions between L7Ae and other RPPs. Our results provide new insights into the architecture and evolution of RNase P, and highlight the utility of native MS and MP in integrated structural biology approaches that seek to augment the information obtained from low/medium-resolution cryo-EM models. | en |
dc.description.notes | NIH [GM-120582 to M.P.F., V.H.W., V.G., P41 GM128577 to V.H.W., S10OD023582 to J.E.J.]; NASA [NNX13AI05G to B.M.]; Virginia Tech Agricultural Experiment Station Hatch Program [CRIS project VA160021 to B.M.]; OSU Comprehensive Cancer Center for a Pelotonia Pre-doctoral Fellowship (to H.D.P.); Virginia Tech Genetics, Bioinformatics, and Computational Biology Doctoral Program Fellowship (to B.K.). Funding for open access charge: University funds. | en |
dc.description.sponsorship | NIH [GM-120582, P41 GM128577, S10OD023582]; NASA [NNX13AI05G]; Virginia Tech Agricultural Experiment Station Hatch Program [CRIS project] [VA160021]; OSU Comprehensive Cancer Center for a Pelotonia Pre-doctoral Fellowship; Virginia Tech Genetics, Bioinformatics, and Computational Biology Doctoral Program Fellowship | en |
dc.description.version | Published version | en |
dc.format.mimetype | application/pdf | en |
dc.identifier.doi | https://doi.org/10.1093/nar/gkac595 | en |
dc.identifier.eissn | 1362-4962 | en |
dc.identifier.issn | 0305-1048 | en |
dc.identifier.issue | 14 | en |
dc.identifier.pmid | 35848927 | en |
dc.identifier.uri | http://hdl.handle.net/10919/112324 | en |
dc.identifier.volume | 50 | en |
dc.language.iso | en | en |
dc.publisher | Oxford University Press | en |
dc.rights | Creative Commons Attribution-NonCommercial 4.0 International | en |
dc.rights.uri | http://creativecommons.org/licenses/by-nc/4.0/ | en |
dc.subject | pyrococcus-horikoshii ot3 | en |
dc.subject | ribonuclease-p | en |
dc.subject | crystal-structure | en |
dc.subject | protein-protein | en |
dc.subject | archaeal homolog | en |
dc.subject | furiosus rpp21 | en |
dc.subject | complex | en |
dc.subject | reconstitution | en |
dc.subject | insights | en |
dc.subject | subunit | en |
dc.title | Elucidation of structure-function relationships in Methanocaldococcus jannaschii RNase P, a multi-subunit catalytic ribonucleoprotein | en |
dc.title.serial | Nucleic Acids Research | en |
dc.type | Article - Refereed | en |
dc.type.dcmitype | Text | en |
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