PXD034224-1

PXD034224 is an original dataset announced via ProteomeXchange.

Title	Supernumerary proteins of the mitochondrial ribosomal small subunit are integral for assembly and translation
Description	Mitochondrial ribosomes (mitoribosomes) have undergone substantial structural remodelling throughout evolution leading to distinct structural properties compared to their prokaryotic ancestors. Compared to their prokaryotic counterparts, mitoribosomes show a substantial loss of ribosomal RNA (rRNA), whilst acquiring unique mitochondrial proteins located on the periphery of the mitoribosome. This extended protein content suggests mitochondrial-specific functions of these proteins involved in mitochondrial mRNA selective-translation activation, ribosome assembly, translation regulation rather than exclusively structural scaffolding. We investigated the functional properties of the 14 unique (mitochondria-specific or supernumerary) mammalian mitoribosomal proteins (snMRP) in the small mitochondrial subunit (mtSSU) by CRISPR-mediated knockout and describe their specific effect on mitoribosome integrity and function. Unexpectedly, we show that each snMRP knockout leads to a unique pattern of mitoribosome instability with variable effects on mitochondrial protein synthesis as essential structural components and all – with the exception of mS37 (MRPS37, CHCHD1) – to almost complete loss of protein synthesis. Concomitantly, through steady-state proteomic analysis we confirm a modular assembly of the mtSSU. Surprisingly, mS37 was found to have impaired stability in all our tested mtSSU and large mitochondrial subunit (mtLSU) snMRPs knockouts as well as in patient-derived cell lines, suggesting an extended role of the protein in mtSSU pre-translation initiation steps. As mitochondrial intermembrane (IMS) targeted protein – a eukaryotic-confined localisation mechanism – with additional unique MIA40 (CHCHD4)-oxidisable motifs involved in the disulfide relay system, mS37 could have a specific role in mitochondrial translation regulation. Indeed, we find oxidation of the cysteines in the CX9C motif of mS37 to be essential for its stability, confirming mS37 regulating mitochondrial translation regulation.
HostingRepository	PRIDE
AnnounceDate	2024-08-09
AnnouncementXML	Submission_2024-08-09_04:30:06.128.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Sachin Singh
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2022-05-30 11:46:50	ID requested
⏵ 1	2024-08-09 04:30:06	announced
2	2024-10-22 06:53:46	announced	2024-10-22: Updated project metadata.

Hilander T, Awadhpersad R, Monteuuis G, Broda KL, Pohjanpelto M, Pyman E, Singh SK, Nyman TA, Crevel I, Taylor RW, Saada A, Balboa D, Battersby BJ, Jackson CB, Carroll CJ, Supernumerary proteins of the human mitochondrial ribosomal small subunit are integral for assembly and translation. iScience, 27(7):110185(2024) [pubmed]

10.1016/j.isci.2024.110185;

submitter keyword: disulfide relay system, IMS, MIA40/CHCHD4, CRISPR/Cas9, mitochondrial disorders, mS37/CHCHD1,mitochondrial ribosome, translation

Christopher J. Carroll
contact affiliation	Genetics Section Molecular and Clinical Sciences St. George's, University of London Jenner 2.142B, Cranmer Terrace London SW17 0RE
contact email	ccarroll@sgul.ac.uk
lab head
Sachin Singh
contact affiliation	Proteomics core facility, Olso University hospital, Norway
contact email	sachin.kumar.singh@rr-research.no
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2024/08/PXD034224

PRIDE project URI

• PRIDE
  1. PXD034224
     1. Label: PRIDE project
     2. Name: Supernumerary proteins of the mitochondrial ribosomal small subunit are integral for assembly and translation