Imaging the fate of histone Cse4 reveals de novo replacement in S phase and subsequent stable residence at centromeres
The budding yeast centromere contains Cse4, a specialized histone H3 variant. Fluorescence pulse-chase analysis of an internally tagged Cse4 reveals that it is replaced with newly synthesized molecules in S phase, remaining stably associated with centromeres thereafter. In contrast, C-terminally-tag...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
eLife Sciences Publications Ltd
2014-05-01
|
| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/02203 |
| _version_ | 1811201047114285056 |
|---|---|
| author | Jan Wisniewski Bassam Hajj Jiji Chen Gaku Mizuguchi Hua Xiao Debbie Wei Maxime Dahan Carl Wu |
| author_facet | Jan Wisniewski Bassam Hajj Jiji Chen Gaku Mizuguchi Hua Xiao Debbie Wei Maxime Dahan Carl Wu |
| author_sort | Jan Wisniewski |
| collection | DOAJ |
| description | The budding yeast centromere contains Cse4, a specialized histone H3 variant. Fluorescence pulse-chase analysis of an internally tagged Cse4 reveals that it is replaced with newly synthesized molecules in S phase, remaining stably associated with centromeres thereafter. In contrast, C-terminally-tagged Cse4 is functionally impaired, showing slow cell growth, cell lethality at elevated temperatures, and extra-centromeric nuclear accumulation. Recent studies using such strains gave conflicting findings regarding the centromeric abundance and cell cycle dynamics of Cse4. Our findings indicate that internally tagged Cse4 is a better reporter of the biology of this histone variant. Furthermore, the size of centromeric Cse4 clusters was precisely mapped with a new 3D-PALM method, revealing substantial compaction during anaphase. Cse4-specific chaperone Scm3 displays steady-state, stoichiometric co-localization with Cse4 at centromeres throughout the cell cycle, while undergoing exchange with a nuclear pool. These findings suggest that a stable Cse4 nucleosome is maintained by dynamic chaperone-in-residence Scm3. |
| first_indexed | 2024-04-12T02:15:16Z |
| format | Article |
| id | doaj.art-14505f5acebb45bc80145b0d78b135ca |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T02:15:16Z |
| publishDate | 2014-05-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-14505f5acebb45bc80145b0d78b135ca2022-12-22T03:52:16ZengeLife Sciences Publications LtdeLife2050-084X2014-05-01310.7554/eLife.02203Imaging the fate of histone Cse4 reveals de novo replacement in S phase and subsequent stable residence at centromeresJan Wisniewski0Bassam Hajj1Jiji Chen2Gaku Mizuguchi3Hua Xiao4Debbie Wei5Maxime Dahan6Carl Wu7Janelia Farm Research Campus, Howard Hughes Medical Institute, Ashburn, United States; Laboratory of Biochemistry and Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, United StatesJanelia Farm Research Campus, Howard Hughes Medical Institute, Ashburn, United StatesJanelia Farm Research Campus, Howard Hughes Medical Institute, Ashburn, United StatesJanelia Farm Research Campus, Howard Hughes Medical Institute, Ashburn, United States; Laboratory of Biochemistry and Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, United StatesLaboratory of Biochemistry and Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, United StatesLaboratory of Biochemistry and Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, United StatesJanelia Farm Research Campus, Howard Hughes Medical Institute, Ashburn, United StatesJanelia Farm Research Campus, Howard Hughes Medical Institute, Ashburn, United States; Laboratory of Biochemistry and Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, United StatesThe budding yeast centromere contains Cse4, a specialized histone H3 variant. Fluorescence pulse-chase analysis of an internally tagged Cse4 reveals that it is replaced with newly synthesized molecules in S phase, remaining stably associated with centromeres thereafter. In contrast, C-terminally-tagged Cse4 is functionally impaired, showing slow cell growth, cell lethality at elevated temperatures, and extra-centromeric nuclear accumulation. Recent studies using such strains gave conflicting findings regarding the centromeric abundance and cell cycle dynamics of Cse4. Our findings indicate that internally tagged Cse4 is a better reporter of the biology of this histone variant. Furthermore, the size of centromeric Cse4 clusters was precisely mapped with a new 3D-PALM method, revealing substantial compaction during anaphase. Cse4-specific chaperone Scm3 displays steady-state, stoichiometric co-localization with Cse4 at centromeres throughout the cell cycle, while undergoing exchange with a nuclear pool. These findings suggest that a stable Cse4 nucleosome is maintained by dynamic chaperone-in-residence Scm3.https://elifesciences.org/articles/02203centromeric nucleosomeCse4 histone variantScm3 chaperonemultifocus microscopyfluorescence pulse-chase3D-PALM |
| spellingShingle | Jan Wisniewski Bassam Hajj Jiji Chen Gaku Mizuguchi Hua Xiao Debbie Wei Maxime Dahan Carl Wu Imaging the fate of histone Cse4 reveals de novo replacement in S phase and subsequent stable residence at centromeres eLife centromeric nucleosome Cse4 histone variant Scm3 chaperone multifocus microscopy fluorescence pulse-chase 3D-PALM |
| title | Imaging the fate of histone Cse4 reveals de novo replacement in S phase and subsequent stable residence at centromeres |
| title_full | Imaging the fate of histone Cse4 reveals de novo replacement in S phase and subsequent stable residence at centromeres |
| title_fullStr | Imaging the fate of histone Cse4 reveals de novo replacement in S phase and subsequent stable residence at centromeres |
| title_full_unstemmed | Imaging the fate of histone Cse4 reveals de novo replacement in S phase and subsequent stable residence at centromeres |
| title_short | Imaging the fate of histone Cse4 reveals de novo replacement in S phase and subsequent stable residence at centromeres |
| title_sort | imaging the fate of histone cse4 reveals de novo replacement in s phase and subsequent stable residence at centromeres |
| topic | centromeric nucleosome Cse4 histone variant Scm3 chaperone multifocus microscopy fluorescence pulse-chase 3D-PALM |
| url | https://elifesciences.org/articles/02203 |
| work_keys_str_mv | AT janwisniewski imagingthefateofhistonecse4revealsdenovoreplacementinsphaseandsubsequentstableresidenceatcentromeres AT bassamhajj imagingthefateofhistonecse4revealsdenovoreplacementinsphaseandsubsequentstableresidenceatcentromeres AT jijichen imagingthefateofhistonecse4revealsdenovoreplacementinsphaseandsubsequentstableresidenceatcentromeres AT gakumizuguchi imagingthefateofhistonecse4revealsdenovoreplacementinsphaseandsubsequentstableresidenceatcentromeres AT huaxiao imagingthefateofhistonecse4revealsdenovoreplacementinsphaseandsubsequentstableresidenceatcentromeres AT debbiewei imagingthefateofhistonecse4revealsdenovoreplacementinsphaseandsubsequentstableresidenceatcentromeres AT maximedahan imagingthefateofhistonecse4revealsdenovoreplacementinsphaseandsubsequentstableresidenceatcentromeres AT carlwu imagingthefateofhistonecse4revealsdenovoreplacementinsphaseandsubsequentstableresidenceatcentromeres |