Mutational and in vitro protein-binding studies on centromere DNA from Saccharomyces cerevisiae - PubMed (original) (raw)
Mutational and in vitro protein-binding studies on centromere DNA from Saccharomyces cerevisiae
R Ng et al. Mol Cell Biol. 1987 Dec.
Abstract
Centromeres on chromosomes in the yeast Saccharomyces cerevisiae contain approximately 140 base pairs (bp) of DNA. The functional centromere (CEN) region contains three important sequence elements (I, PuTCACPuTG; II, 78 to 86 bp of high-AT DNA; and III, a conserved 25-bp sequence with internal bilateral symmetry). Various point mutations or deletions in the element III region have a profound effect on CEN function in vivo, indicating that this DNA region is a key protein-binding site. This has been confirmed by the use of two in vitro assays to detect binding of yeast proteins to DNA fragments containing wild-type or mutationally altered CEN3 sequences. An exonuclease III protection assay was used to demonstrate specific binding of proteins to the element III region of CEN3. In addition, a gel DNA fragment mobility shift assay was used to characterize the binding reaction parameters. Sequence element III mutations that inactivate CEN function in vivo also prevent binding of proteins in the in vitro assays. The mobility shift assay indicates that double-stranded DNAs containing sequence element III efficiently bind proteins in the absence of sequence elements I and II, although the latter sequences are essential for optimal CEN function in vivo.
Similar articles
- Structural studies on centromeres in the yeast Saccharomyces cerevisiae.
Ng R, Ness J, Carbon J. Ng R, et al. Basic Life Sci. 1986;40:479-92. doi: 10.1007/978-1-4684-5251-8_36. Basic Life Sci. 1986. PMID: 3032143 - Mutational analysis of meiotic and mitotic centromere function in Saccharomyces cerevisiae.
Cumberledge S, Carbon J. Cumberledge S, et al. Genetics. 1987 Oct;117(2):203-12. doi: 10.1093/genetics/117.2.203. Genetics. 1987. PMID: 3311877 Free PMC article. - The yeast centromere CDEI/Cpf1 complex: differences between in vitro binding and in vivo function.
Wilmen A, Pick H, Niedenthal RK, Sen-Gupta M, Hegemann JH. Wilmen A, et al. Nucleic Acids Res. 1994 Jul 25;22(14):2791-800. doi: 10.1093/nar/22.14.2791. Nucleic Acids Res. 1994. PMID: 8052535 Free PMC article. - The structure and function of yeast centromeres.
Clarke L, Carbon J. Clarke L, et al. Annu Rev Genet. 1985;19:29-55. doi: 10.1146/annurev.ge.19.120185.000333. Annu Rev Genet. 1985. PMID: 3909945 Review. No abstract available. - Centromeres of budding and fission yeasts.
Clarke L. Clarke L. Trends Genet. 1990 May;6(5):150-4. doi: 10.1016/0168-9525(90)90149-z. Trends Genet. 1990. PMID: 2195725 Review.
Cited by
- Cryo-EM structure of the complete inner kinetochore of the budding yeast point centromere.
Dendooven T, Zhang Z, Yang J, McLaughlin SH, Schwab J, Scheres SHW, Yatskevich S, Barford D. Dendooven T, et al. Sci Adv. 2023 Jul 28;9(30):eadg7480. doi: 10.1126/sciadv.adg7480. Epub 2023 Jul 28. Sci Adv. 2023. PMID: 37506202 Free PMC article. - Nanoscale structural organization and stoichiometry of the budding yeast kinetochore.
Cieslinski K, Wu YL, Nechyporenko L, Hörner SJ, Conti D, Skruzny M, Ries J. Cieslinski K, et al. J Cell Biol. 2023 Apr 3;222(4):e202209094. doi: 10.1083/jcb.202209094. Epub 2023 Jan 27. J Cell Biol. 2023. PMID: 36705601 Free PMC article. - A transcriptional roadblock protects yeast centromeres.
Hedouin S, Logsdon GA, Underwood JG, Biggins S. Hedouin S, et al. Nucleic Acids Res. 2022 Aug 12;50(14):7801-7815. doi: 10.1093/nar/gkac117. Nucleic Acids Res. 2022. PMID: 35253883 Free PMC article. - Identification and characterization of centromeric sequences in Xenopus laevis.
Smith OK, Limouse C, Fryer KA, Teran NA, Sundararajan K, Heald R, Straight AF. Smith OK, et al. Genome Res. 2021 Jun;31(6):958-967. doi: 10.1101/gr.267781.120. Epub 2021 Apr 19. Genome Res. 2021. PMID: 33875480 Free PMC article. - The COMA complex interacts with Cse4 and positions Sli15/Ipl1 at the budding yeast inner kinetochore.
Fischböck-Halwachs J, Singh S, Potocnjak M, Hagemann G, Solis-Mezarino V, Woike S, Ghodgaonkar-Steger M, Weissmann F, Gallego LD, Rojas J, Andreani J, Köhler A, Herzog F. Fischböck-Halwachs J, et al. Elife. 2019 May 21;8:e42879. doi: 10.7554/eLife.42879. Elife. 2019. PMID: 31112132 Free PMC article.
References
- J Cell Biol. 1987 Jan;104(1):9-18 - PubMed
- Mol Cell Biol. 1987 Jan;7(1):403-9 - PubMed
- Mol Cell Biol. 1985 Nov;5(11):2887-93 - PubMed
- Proc Natl Acad Sci U S A. 1979 Mar;76(3):1035-9 - PubMed
- Cell. 1984 Jun;37(2):351-3 - PubMed
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Molecular Biology Databases