The nuclear location and chromatin organization of active chorion amplification origins

B. R. Calvi, A. C. Spradling

Research output: Contribution to journalArticle

Abstract

It remains unclear how certain regions on metazoan chromosomes are selected to initiate DNA replication. In recent years a number of origins of DNA replication have been mapped, but there is still no DNA consensus for predicting where replication will initiate. Evidence suggests that the higher order structure of the nucleus and chromosome influences origin activity. Chromosomal DNA replication is proposed to occur in special compartments in the nucleus called replication foci. Foci in different regions of the nucleus initiate replication at different times of S-phase, suggesting nuclear position may contribute to where and when replication begins. Here we test the contribution of nuclear compartments for well-defined origins, those involved in amplification of the chorion (eggshell) genes during Drosophila oogenesis. The results of three-dimensional confocal microscopy indicate that chorion DNA replication origins are highly active in diverse positions within the nucleus. We also find that chorion replication origins inserted at ectopic chromosomal sites can amplify highly in diverse nuclear locations distinct from the endogenous loci, including when they are buffered against genomic position effects. We used fluorescence in situ hybridization to analyze chromosome structure during amplification. Contrary to the replication factory model, we find no evidence for spooling of DNA toward a replication center. We discuss the implications of these results for understanding the role of higher order structure in amplification and chromosome duplication.

Original languageEnglish (US)
Pages (from-to)159-172
Number of pages14
JournalChromosoma
Volume110
Issue number3
StatePublished - 2001
Externally publishedYes

Fingerprint

Chorion
DNA Replication
Replication Origin
Chromatin
Chromosome Structures
Chromosome Duplication
Egg Shell
Oogenesis
DNA
Fluorescence In Situ Hybridization
S Phase
Confocal Microscopy
Drosophila
Chromosomes
Genes

ASJC Scopus subject areas

  • Genetics

Cite this

The nuclear location and chromatin organization of active chorion amplification origins. / Calvi, B. R.; Spradling, A. C.

In: Chromosoma, Vol. 110, No. 3, 2001, p. 159-172.

Research output: Contribution to journalArticle

Calvi, BR & Spradling, AC 2001, 'The nuclear location and chromatin organization of active chorion amplification origins', Chromosoma, vol. 110, no. 3, pp. 159-172.
Calvi, B. R. ; Spradling, A. C. / The nuclear location and chromatin organization of active chorion amplification origins. In: Chromosoma. 2001 ; Vol. 110, No. 3. pp. 159-172.
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