Pietrasanta, Lia I. and Thrower, Douglas and Hsieh, Wan and Rao, Shashirekha and Stemmann, Olaf and Lechner, Johannes and Carbon, John and Hansma, Helen (1999) Probing the Saccharomyces cerevisiae centromeric DNA (CEN DNA)-binding factor 3 (CBF3) kinetochore complex by using atomic force microscopy. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 96 (7). pp. 3757-3762. ISSN 0027-8424,
Full text not available from this repository. (Request a copy)Abstract
Yeast centromeric DNA (CEN DNA) binding factor 3 (CBF3) is a multisubunit protein complex that binds to the essential CDEIII element in CEN DNA. The four CBF3 proteins are required for accurate chromosome segregation and are considered to be core components of the yeast kinetochore. We have examined the structure of the CBF3-CEN DNA complex by atomic force microscopy, Assembly of CBF3-CEN DNA complexes was performed by combining purified CBF3 proteins with a DNA fragment that includes the CEN region from yeast chromosome III. Atomic force microscopy images showed DNA molecules with attached globular bodies. The contour length of the DNA containing the complex is approximate to 9% shorter than the DNA alone, suggesting some winding of DNA within the complex, The measured location of the single binding site indicates that the complex is located asymmetrically to the right of CDEIII extending away from CDEI and CDEII, which is consistent with previous data. The CEN DNA is bent approximate to 55 degrees at the site of complex formation. A significant fraction of the complexes are linked in pairs, showing three to four DNA arms,,vith molecular volumes approximately three times the mean volumes of two-armed complexes, These multi-armed complexes indicate that CBF3 can bind two DNA molecules together in vitro and, thus, may be involved in holding together chromatid pairs during mitosis.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | DEPENDENT PROTEIN-KINASE; CRYSTAL-STRUCTURE; BINDING; RECOGNITION; SPECIFICITY; COMPONENT; GENE; AIR; KU; |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 08 Nov 2022 15:07 |
| Last Modified: | 08 Nov 2022 15:07 |
| URI: | https://pred.uni-regensburg.de/id/eprint/48421 |
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