A Reversible Association between Smc Coiled Coils Is Regulated by Lysine Acetylation and Is Required for Cohesin Association with the DNA.
Cohesin is a ring-shaped protein complex that is capable of embracing DNA. Most of the ring circumference is comprised of the anti-parallel intramolecular coiled coils of the Smc1 and Smc3 proteins, which connect globular head and hinge domains. Smc coiled coil arms contain multiple acetylated and ubiquitylated lysines. To investigate the role of these modifications, we substituted lysines for arginines to mimic the unmodified state and uncovered genetic interaction between the Smc arms. Using scanning force microscopy, we show that wild-type Smc arms associate with each other when the complex is not on DNA. Deacetylation of the Smc1/Smc3 dimers promotes arms' dissociation. Smc arginine mutants display loose packing of the Smc arms and, although they dimerize at the hinges, fail to connect the heads and associate with the DNA. Our findings highlight the importance of a "collapsed ring," or "rod," conformation of cohesin for its loading on the chromosomes.
Bioinformatics Institute, A(∗)STAR, Singapore 138671, Singapore; Friedrich Miescher Laboratory of the Max Planck Society, Tuebingen 72076, Germany.
Irina
I
Kulemzina
Bioinformatics Institute, A(∗)STAR, Singapore 138671, Singapore.
Keven
K
Ang
Mechanobiology Institute, National University of Singapore, Singapore 117411, Singapore.
Xiaodan
X
Zhao
Bioinformatics Institute, A(∗)STAR, Singapore 138671, Singapore.
Jun-Thing
JT
Teh
Friedrich Miescher Laboratory of the Max Planck Society, Tuebingen 72076, Germany.
Vikash
V
Verma
Bioinformatics Institute, A(∗)STAR, Singapore 138671, Singapore.
Sasikala
S
Suranthran
Bioinformatics Institute, A(∗)STAR, Singapore 138671, Singapore.
Alap P
AP
Chavda
Bioinformatics Institute, A(∗)STAR, Singapore 138671, Singapore.
Roland G
RG
Huber
Bioinformatics Institute, A(∗)STAR, Singapore 138671, Singapore.
Birgit
B
Eisenhaber
Bioinformatics Institute, A(∗)STAR, Singapore 138671, Singapore; School of Computer Engineering, Nanyang Technological University, Singapore 637553, Singapore; Department of Biological Sciences, National University of Singapore, Singapore 117597, Singapore.
Frank
F
Eisenhaber
Mechanobiology Institute, National University of Singapore, Singapore 117411, Singapore; Department of Physics, National University of Singapore, Singapore 117551, Singapore; Center for Bioimaging Sciences, National University of Singapore, Singapore 117557, Singapore.
Jie
J
Yan
Bioinformatics Institute, A(∗)STAR, Singapore 138671, Singapore; Institute of Molecular and Cell Biology, A(∗)STAR, Singapore 138673, Singapore; Friedrich Miescher Laboratory of the Max Planck Society, Tuebingen 72076, Germany; Department of Physics, National University of Singapore, Singapore 117551, Singapore. Electronic address: dmitryi@bii.a-star.edu.sg.
Dmitri
D
Ivanov
D000107:Acetylation; D019943:Amino Acid Substitution; D000818:Animals; D001120:Arginine / Q000378:metabolism; D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002842:Chromatids / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D015825:Chromosomes, Fungal / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D015966:Gene Expression Regulation, Fungal; D008239:Lysine / Q000378:metabolism*; D000072756:Protein Conformation, alpha-Helical; D054730:Protein Interaction Domains and Motifs; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D061987:Sf9 Cells; D015398:Signal Transduction; D018411:Spodoptera; D000097722:Cohesins; D000098550:Structural Maintenance of Chromosome Protein 1
D001120:Arginine; D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; D004271:DNA, Fungal; D008239:Lysine; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D000097722:Cohesins; D000098550:Structural Maintenance of Chromosome Protein 1; C120372:SMC3 protein, S cerevisiae