A Meier-Gorlin Syndrome Mutation in a Conserved C-terminal Helix of Orc6 Impedes Origin Recognition Complex Formation

Overview

Journal Elife

Specialty Biology

Date 2013 Oct 19

PMID 24137536

Citations 39

Authors

Franziska Bleichert

Maxim Balasov

Igor Chesnokov

Eva Nogales

Michael R Botchan

James M Berger

Affiliations

Soon will be listed here.

Abstract

In eukaryotes, DNA replication requires the origin recognition complex (ORC), a six-subunit assembly that promotes replisome formation on chromosomal origins. Despite extant homology between certain subunits, the degree of structural and organizational overlap between budding yeast and metazoan ORC has been unclear. Using 3D electron microscopy, we determined the subunit organization of metazoan ORC, revealing that it adopts a global architecture very similar to the budding yeast complex. Bioinformatic analysis extends this conservation to Orc6, a subunit of somewhat enigmatic function. Unexpectedly, a mutation in the Orc6 C-terminus linked to Meier-Gorlin syndrome, a dwarfism disorder, impedes proper recruitment of Orc6 into ORC; biochemical studies reveal that this region of Orc6 associates with a previously uncharacterized domain of Orc3 and is required for ORC function and MCM2-7 loading in vivo. Together, our results suggest that Meier-Gorlin syndrome mutations in Orc6 impair the formation of ORC hexamers, interfering with appropriate ORC functions. DOI:http://dx.doi.org/10.7554/eLife.00882.001.

Citing Articles

MCM2-7 ring closure involves the Mcm5 C-terminus and triggers Mcm4 ATP hydrolysis.

Faull S, Barbon M, Mossler A, Yuan Z, Bai L, Reuter L Nat Commun. 2025; 16(1):14.

PMID: 39747125 PMC: 11695723. DOI: 10.1038/s41467-024-55479-1.

MCM double hexamer loading visualized with human proteins.

Weissmann F, Greiwe J, Puhringer T, Eastwood E, Couves E, Miller T Nature. 2024; 636(8042):499-508.

PMID: 39604733 PMC: 11634765. DOI: 10.1038/s41586-024-08263-6.

Multiple mechanisms for licensing human replication origins.

Yang R, Hunker O, Wise M, Bleichert F Nature. 2024; 636(8042):488-498.

PMID: 39604729 DOI: 10.1038/s41586-024-08237-8.

Multiple pathways for licensing human replication origins.

Yang R, Hunker O, Wise M, Bleichert F bioRxiv. 2024; .

PMID: 38645015 PMC: 11030351. DOI: 10.1101/2024.04.10.588796.

Ordered and disordered regions of the Origin Recognition Complex direct differential in vivo binding at distinct motif sequences.

Chappleboim M, Naveh-Tassa S, Carmi M, Levy Y, Barkai N Nucleic Acids Res. 2024; 52(10):5720-5731.

PMID: 38597680 PMC: 11162778. DOI: 10.1093/nar/gkae249.

References

Kneissl M, Putter V, Szalay A, Grummt F . Interaction and assembly of murine pre-replicative complex proteins in yeast and mouse cells. J Mol Biol. 2003; 327(1):111-28. DOI: 10.1016/s0022-2836(03)00079-2. View

Lee D, Bell S . Architecture of the yeast origin recognition complex bound to origins of DNA replication. Mol Cell Biol. 1997; 17(12):7159-68. PMC: 232573. DOI: 10.1128/MCB.17.12.7159. View

Vashee S, Simancek P, Challberg M, Kelly T . Assembly of the human origin recognition complex. J Biol Chem. 2001; 276(28):26666-73. DOI: 10.1074/jbc.M102493200. View

de Munnik S, Bicknell L, Aftimos S, Al-Aama J, van Bever Y, Bober M . Meier-Gorlin syndrome genotype-phenotype studies: 35 individuals with pre-replication complex gene mutations and 10 without molecular diagnosis. Eur J Hum Genet. 2012; 20(6):598-606. PMC: 3355263. DOI: 10.1038/ejhg.2011.269. View

Frigola J, Remus D, Mehanna A, Diffley J . ATPase-dependent quality control of DNA replication origin licensing. Nature. 2013; 495(7441):339-43. PMC: 4825857. DOI: 10.1038/nature11920. View

Bicknell L, Walker S, Klingseisen A, Stiff T, Leitch A, Kerzendorfer C . Mutations in ORC1, encoding the largest subunit of the origin recognition complex, cause microcephalic primordial dwarfism resembling Meier-Gorlin syndrome. Nat Genet. 2011; 43(4):350-5. DOI: 10.1038/ng.776. View

Nikolov D, Chen H, Halay E, Usheva A, Hisatake K, Lee D . Crystal structure of a TFIIB-TBP-TATA-element ternary complex. Nature. 1995; 377(6545):119-28. DOI: 10.1038/377119a0. View

Sorzano C, Marabini R, Velazquez-Muriel J, Bilbao-Castro J, Scheres S, Carazo J . XMIPP: a new generation of an open-source image processing package for electron microscopy. J Struct Biol. 2004; 148(2):194-204. DOI: 10.1016/j.jsb.2004.06.006. View

Abbas T, Keaton M, Dutta A . Genomic instability in cancer. Cold Spring Harb Perspect Biol. 2013; 5(3):a012914. PMC: 3578360. DOI: 10.1101/cshperspect.a012914. View

10.

Tang G, Peng L, Baldwin P, Mann D, Jiang W, Rees I . EMAN2: an extensible image processing suite for electron microscopy. J Struct Biol. 2006; 157(1):38-46. DOI: 10.1016/j.jsb.2006.05.009. View

11.

Iyer L, Leipe D, Koonin E, Aravind L . Evolutionary history and higher order classification of AAA+ ATPases. J Struct Biol. 2004; 146(1-2):11-31. DOI: 10.1016/j.jsb.2003.10.010. View

12.

Lander G, Stagg S, Voss N, Cheng A, Fellmann D, Pulokas J . Appion: an integrated, database-driven pipeline to facilitate EM image processing. J Struct Biol. 2009; 166(1):95-102. PMC: 2775544. DOI: 10.1016/j.jsb.2009.01.002. View

13.

Ludtke S, Baldwin P, Chiu W . EMAN: semiautomated software for high-resolution single-particle reconstructions. J Struct Biol. 1999; 128(1):82-97. DOI: 10.1006/jsbi.1999.4174. View

14.

Dueber E, Corn J, Bell S, Berger J . Replication origin recognition and deformation by a heterodimeric archaeal Orc1 complex. Science. 2007; 317(5842):1210-3. DOI: 10.1126/science.1143690. View

15.

Neuwald A, Aravind L, Spouge J, Koonin E . AAA+: A class of chaperone-like ATPases associated with the assembly, operation, and disassembly of protein complexes. Genome Res. 1999; 9(1):27-43. View

16.

Balasov M, Huijbregts R, Chesnokov I . Role of the Orc6 protein in origin recognition complex-dependent DNA binding and replication in Drosophila melanogaster. Mol Cell Biol. 2007; 27(8):3143-53. PMC: 1899928. DOI: 10.1128/MCB.02382-06. View

17.

Siddiqui K, Stillman B . ATP-dependent assembly of the human origin recognition complex. J Biol Chem. 2007; 282(44):32370-83. DOI: 10.1074/jbc.M705905200. View

18.

Stiff T, Alagoz M, Alcantara D, Outwin E, Brunner H, Bongers E . Deficiency in origin licensing proteins impairs cilia formation: implications for the aetiology of Meier-Gorlin syndrome. PLoS Genet. 2013; 9(3):e1003360. PMC: 3597520. DOI: 10.1371/journal.pgen.1003360. View

19.

Randell J, Bowers J, Rodriguez H, Bell S . Sequential ATP hydrolysis by Cdc6 and ORC directs loading of the Mcm2-7 helicase. Mol Cell. 2006; 21(1):29-39. DOI: 10.1016/j.molcel.2005.11.023. View

20.

Chesnokov I, Gossen M, Remus D, Botchan M . Assembly of functionally active Drosophila origin recognition complex from recombinant proteins. Genes Dev. 1999; 13(10):1289-96. PMC: 316721. DOI: 10.1101/gad.13.10.1289. View