Construction and Analysis of Mouse Strains Lacking the Ubiquitin Ligase UBR1 (E3alpha) of the N-end Rule Pathway

Overview

Journal Mol Cell Biol

Specialty Cell Biology

Date 2001 Nov 2

PMID 11689692

Citations 62

Authors

Y T Kwon

Z Xia

I V Davydov

S H Lecker

A Varshavsky

Affiliations

Soon will be listed here.

Abstract

The N-end rule relates the in vivo half-life of a protein to the identity of its N-terminal residue. In the yeast Saccharomyces cerevisiae, the UBR1-encoded ubiquitin ligase (E3) of the N-end rule pathway mediates the targeting of substrate proteins in part through binding to their destabilizing N-terminal residues. The functions of the yeast N-end rule pathway include fidelity of chromosome segregation and the regulation of peptide import. Our previous work described the cloning of cDNA and a gene encoding the 200-kDa mouse UBR1 (E3alpha). Here we show that mouse UBR1, in the presence of a cognate mouse ubiquitin-conjugating (E2) enzyme, can rescue the N-end rule pathway in ubr1Delta S. cerevisiae. We also constructed UBR1(-/-) mouse strains that lacked the UBR1 protein. UBR1(-/-) mice were viable and fertile but weighed significantly less than congenic +/+ mice. The decreased mass of UBR1(-/-) mice stemmed at least in part from smaller amounts of the skeletal muscle and adipose tissues. The skeletal muscle of UBR1(-/-) mice apparently lacked the N-end rule pathway and exhibited abnormal regulation of fatty acid synthase upon starvation. By contrast, and despite the absence of the UBR1 protein, UBR1(-/-) fibroblasts contained the N-end rule pathway. Thus, UBR1(-/-) mice are mosaics in regard to the activity of this pathway, owing to differential expression of proteins that can substitute for the ubiquitin ligase UBR1 (E3alpha). We consider these UBR1-like proteins and discuss the functions of the mammalian N-end rule pathway.

Citing Articles

Metabolic regulation of proteome stability via N-terminal acetylation controls male germline stem cell differentiation and reproduction.

Francois C, Pihl T, Dunoyer de Segonzac M, Herault C, Hudry B Nat Commun. 2023; 14(1):6737.

PMID: 37872135 PMC: 10593830. DOI: 10.1038/s41467-023-42496-9.

UBR-1 ubiquitin ligase regulates the balance between GABAergic and glutamatergic signaling.

Li Y, Chitturi J, Yu B, Zhang Y, Wu J, Ti P EMBO Rep. 2023; 24(11):e57014.

PMID: 37811674 PMC: 10626437. DOI: 10.15252/embr.202357014.

Targeted degradation of ⍺-synuclein aggregates in Parkinson's disease using the AUTOTAC technology.

Lee J, Sung K, Bae E, Yoon D, Kim D, Lee J Mol Neurodegener. 2023; 18(1):41.

PMID: 37355598 PMC: 10290391. DOI: 10.1186/s13024-023-00630-7.

Transferase-Mediated Labeling of Protein N-Termini with Click Chemistry Handles.

Wagner A, Warner J, Garrett H, Walters C, Petersson E Methods Mol Biol. 2023; 2620:157-175.

PMID: 37010762 DOI: 10.1007/978-1-0716-2942-0_21.

N-end Rule-Mediated Proteasomal Degradation of ATGL Promotes Lipid Storage.

Xu J, Liu Z, Zhang J, Chen S, Wang W, Zhao X Diabetes. 2022; 72(2):210-222.

PMID: 36346641 PMC: 9871197. DOI: 10.2337/db22-0362.

References

Li J, Pickart C . Binding of phenylarsenoxide to Arg-tRNA protein transferase is independent of vicinal thiols. Biochemistry. 1995; 34(48):15829-37. DOI: 10.1021/bi00048a028. View

Stewart A, ARFIN S, Bradshaw R . The sequence of porcine protein NH2-terminal asparagine amidohydrolase. A new component of the N-end Rule pathway. J Biol Chem. 1995; 270(1):25-8. DOI: 10.1074/jbc.270.1.25. View

Rao H, Uhlmann F, Nasmyth K, Varshavsky A . Degradation of a cohesin subunit by the N-end rule pathway is essential for chromosome stability. Nature. 2001; 410(6831):955-9. DOI: 10.1038/35073627. View

Baker R, Varshavsky A . Inhibition of the N-end rule pathway in living cells. Proc Natl Acad Sci U S A. 1991; 88(4):1090-4. PMC: 50962. DOI: 10.1073/pnas.88.4.1090. View

Alagramam K, Naider F, Becker J . A recognition component of the ubiquitin system is required for peptide transport in Saccharomyces cerevisiae. Mol Microbiol. 1995; 15(2):225-34. DOI: 10.1111/j.1365-2958.1995.tb02237.x. View

Schauber C, Chen L, Tongaonkar P, Vega I, Madura K . Sequence elements that contribute to the degradation of yeast G alpha. Genes Cells. 1998; 3(5):307-19. DOI: 10.1046/j.1365-2443.1998.00192.x. View

Balzi E, Choder M, Chen W, Varshavsky A, Goffeau A . Cloning and functional analysis of the arginyl-tRNA-protein transferase gene ATE1 of Saccharomyces cerevisiae. J Biol Chem. 1990; 265(13):7464-71. View

Hershko A, Ciechanover A . The ubiquitin system. Annu Rev Biochem. 1998; 67:425-79. DOI: 10.1146/annurev.biochem.67.1.425. View

Madura K, Varshavsky A . Degradation of G alpha by the N-end rule pathway. Science. 1994; 265(5177):1454-8. DOI: 10.1126/science.8073290. View

10.

Hicke L . Protein regulation by monoubiquitin. Nat Rev Mol Cell Biol. 2001; 2(3):195-201. DOI: 10.1038/35056583. View

11.

Suzuki T, Varshavsky A . Degradation signals in the lysine-asparagine sequence space. EMBO J. 1999; 18(21):6017-26. PMC: 1171667. DOI: 10.1093/emboj/18.21.6017. View

12.

Bachmair A, Finley D, Varshavsky A . In vivo half-life of a protein is a function of its amino-terminal residue. Science. 1986; 234(4773):179-86. DOI: 10.1126/science.3018930. View

13.

Tyers M, Jorgensen P . Proteolysis and the cell cycle: with this RING I do thee destroy. Curr Opin Genet Dev. 2000; 10(1):54-64. DOI: 10.1016/s0959-437x(99)00049-0. View

14.

TURNER G, Varshavsky A . Detecting and measuring cotranslational protein degradation in vivo. Science. 2000; 289(5487):2117-20. DOI: 10.1126/science.289.5487.2117. View

15.

Kwon Y, Reiss Y, Fried V, Hershko A, Yoon J, Gonda D . The mouse and human genes encoding the recognition component of the N-end rule pathway. Proc Natl Acad Sci U S A. 1998; 95(14):7898-903. PMC: 20901. DOI: 10.1073/pnas.95.14.7898. View

16.

Plemper R, Wolf D . Retrograde protein translocation: ERADication of secretory proteins in health and disease. Trends Biochem Sci. 1999; 24(7):266-70. DOI: 10.1016/s0968-0004(99)01420-6. View

17.

Xie Y, Varshavsky A . The E2-E3 interaction in the N-end rule pathway: the RING-H2 finger of E3 is required for the synthesis of multiubiquitin chain. EMBO J. 1999; 18(23):6832-44. PMC: 1171746. DOI: 10.1093/emboj/18.23.6832. View

18.

Waizenegger I, Hauf S, Meinke A, Peters J . Two distinct pathways remove mammalian cohesin from chromosome arms in prophase and from centromeres in anaphase. Cell. 2000; 103(3):399-410. DOI: 10.1016/s0092-8674(00)00132-x. View

19.

Sijts A, Pilip I, Pamer E . The Listeria monocytogenes-secreted p60 protein is an N-end rule substrate in the cytosol of infected cells. Implications for major histocompatibility complex class I antigen processing of bacterial proteins. J Biol Chem. 1997; 272(31):19261-8. DOI: 10.1074/jbc.272.31.19261. View

20.

Laney J, Hochstrasser M . Substrate targeting in the ubiquitin system. Cell. 1999; 97(4):427-30. DOI: 10.1016/s0092-8674(00)80752-7. View