Formyl-methionine as an N-degron of a eukaryotic N-end rule pathway

Kim, Jeong-Mok and Seok, Ok-Hee and Ju, Shinyeong and Heo, Ji-Eun and Yeom, Jeonghun and Kim, Da-Som and Yoo, Joo-Yeon and Varshavsky, Alexander and Lee, Cheolju and Hwang, Cheol-Sang (2018) Formyl-methionine as an N-degron of a eukaryotic N-end rule pathway. Science, 362 (6418). Art. No. eaat0174. ISSN 0036-8075. PMCID PMC6551516. doi:10.1126/science.aat0174. https://resolver.caltech.edu/CaltechAUTHORS:20181108-125308990

	PDF - Accepted Version See Usage Policy. 1MB
	PDF (Materials and Methods; Figs. S1 to S12; Tables S1 to S5; References) - Supplemental Material See Usage Policy. 5MB

Use this Persistent URL to link to this item: https://resolver.caltech.edu/CaltechAUTHORS:20181108-125308990

Abstract

In bacteria, nascent proteins bear the pretranslationally generated N-terminal (Nt) formyl-methionine (fMet) residue. Nt-fMet of bacterial proteins is a degradation signal, termed fMet/N-degron. In contrast, proteins synthesized by cytosolic ribosomes of eukaryotes were presumed to bear unformylated Nt-Met. Here we found that the yeast formyltransferase Fmt1, although imported into mitochondria, could also produce Nt-formylated proteins in the cytosol. Nt-formylated proteins were strongly up-regulated in stationary phase or upon starvation for specific amino acids. This up-regulation strictly required the Gcn2 kinase, which phosphorylates Fmt1 and mediates its retention in the cytosol. We also found that the Nt-fMet residues of Nt-formylated proteins act as fMet/N-degrons, and identified the Psh1 ubiquitin ligase as the recognition component of this eukaryotic fMet/N-end rule pathway, which destroys Nt-formylated proteins.

Item Type:

Article

Related URLs:

URL	URL Type	Description
https://doi.org/10.1126/science.aat0174	DOI	Article
https://www.sciencemag.org/cgi/content/full/science.aat0174/DC1	Publisher	Supplementary Materials
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6551516	PubMed Central	Article

ORCID:

Author	ORCID
Kim, Jeong-Mok	0000-0002-7223-248X
Ju, Shinyeong	0000-0001-5483-4690
Yoo, Joo-Yeon	0000-0001-5942-3989
Varshavsky, Alexander	0000-0002-4011-258X
Lee, Cheolju	0000-0001-8482-4696
Hwang, Cheol-Sang	0000-0002-0105-5957

Additional Information:

© 2018 American Association for the Advancement of Science. We thank W.-K. Huh (Seoul National University, Seoul, Korea) for yeast deletion library mutants, S. Biggins (Hutchinson Cancer Center, Seattle, WA, USA) for pSB1535 and pSB1541, and R. Sauer (MIT, Cambridge, MA, USA) for AG110A(DE) E. coli. We also thank the present and former members of the Hwang laboratory for their assistance and advice. Supported by grants from the Samsung Science & Technology Foundation (SSTF-BA1401-17) and the BK21 plus program (C.-S.H.), by the NRF grants of the Korean Government (MSIP) NRF-2017M3A9F9030559 (C.L.) and NRF-2017R1A5A1015366 (J.-Y.Y.), and by NIH grants R01GM031530 and R01DK039520 (A.V.). Author contributions: C.-S.H., J.-M.K., J.-Y.Y., C.L., A.V., and other coauthors designed research. J.-M.K., O.-H.S., S.J., J.-E.H., J.Y., D.-S.K. and C.-S.H. performed research, and all coauthors analyzed data. C.-S.H., J.-M.K., C.L. and A.V. wrote the paper. All coauthors declare no competing interests. Data and materials availability: All mass spectrometric data of this study are available in PRIDE database (accession number: PXD010780). All (other) data needed to evaluate the conclusions in the paper are present in the paper or the Supplementary Materials.

Funders:

Funding Agency	Grant Number
Samsung Science & Technology Foundation	SSTF-BA1401-17
Korean Government Project	BK21
National Research Foundation of Korea	NRF-2017M3A9F9030559
National Research Foundation of Korea	NRF-2017R1A5A1015366
NIH	R01GM031530
NIH	R01DK039520

Issue or Number:

6418

PubMed Central ID:

PMC6551516

DOI:

10.1126/science.aat0174

Record Number:

CaltechAUTHORS:20181108-125308990

Persistent URL:

https://resolver.caltech.edu/CaltechAUTHORS:20181108-125308990

Official Citation:

Formyl-methionine as an N-degron of a eukaryotic N-end rule pathway. BY JEONG-MOK KIM, OK-HEE SEOK, SHINYEONG JU, JI-EUN HEO, JEONGHUN YEOM, DA-SOM KIM, JOO-YEON YOO, ALEXANDER VARSHAVSKY, CHEOLJU LEE, CHEOL-SANG HWANG. SCIENCE 30 NOV 2018

Usage Policy:

No commercial reproduction, distribution, display or performance rights in this work are provided.

ID Code:

90756

Collection:

CaltechAUTHORS

Deposited By:

Tony Diaz

Deposited On:

08 Nov 2018 21:03

Last Modified:

16 Nov 2021 03:35

Repository Staff Only: item control page