Antisense knockdown of cyclin E does not affect the midblastula transition in Xenopus laevis embryos

Michael K Slevin; Gillian Lyons-Levy; Daniel L Weeks; Rebecca S Hartley

doi:10.4161/cc.4.10.2035

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Antisense knockdown of cyclin E does not affect the midblastula transition in Xenopus laevis embryos

Journal article

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Antisense knockdown of cyclin E does not affect the midblastula transition in Xenopus laevis embryos

Michael K Slevin, Gillian Lyons-Levy, Daniel L Weeks and Rebecca S Hartley

Cell cycle (Georgetown, Tex.), Vol.4(10), pp.1396-1402

10/2005

DOI: 10.4161/cc.4.10.2035

PMID: 16131839

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Abstract

In Xenopus laevis embryos, cyclin E protein remains constitutively high throughout the first 12 cell cycles following fertilization until the onset of the midblastula transition (MBT) (after the 12(th) cell cycle) when it undergoes a dramatic reduction. The disappearance of cyclin E at the MBT occurs independently of active cell cycle progression, zygotic transcription, protein synthesis and the nuclear to cytoplasmic ratio. This has suggested that cyclin E is part of an autonomous maternal timer that regulates the onset of the MBT. To determine how constitutively high levels of cyclin E are maintained prior to the MBT and to investigate if the reduction in cyclin E protein affects the timing of the MBT, we have knocked down endogenous cyclin E mRNA using an N,N-diethyl-ethylene-diamine modified antisense oligonucleotide targeted to its open reading frame. We report that maintenance of high levels of cyclin E protein before the MBT is due to a balance between ongoing translation and proteolytic degradation. In support of our antisense experiments, polysome analysis demonstrates that cyclin E mRNA is associated with the translated fraction prior to the MBT. Furthermore, knockdown of cyclin E was not associated with defects in the timing of developmental events. Our data suggests that cyclin E is not required for the later cell cycles of embryonic development and that the pathway effecting downregulation of cyclin E rather then cyclin E degradation itself may be part of a maternal timer that affects the onset of the MBT.

Cyclin E - deficiency

Xenopus laevis - embryology

Cyclin A - metabolism

RNA, Messenger - genetics

Embryo, Nonmammalian - metabolism

Embryo, Nonmammalian - embryology

Oligodeoxyribonucleotides, Antisense - genetics

Cyclin A - genetics

Cyclin E - genetics

Animals

Xenopus laevis - metabolism

Base Sequence

Polyribosomes - metabolism

Protein Biosynthesis - genetics

Xenopus laevis - genetics

Cyclin E - metabolism

Oligodeoxyribonucleotides, Antisense - metabolism

Details

Title: Subtitle: Antisense knockdown of cyclin E does not affect the midblastula transition in Xenopus laevis embryos
Creators: Michael K Slevin - Molecular and Cellular Biology Program, University of Iowa, Iowa City, Iowa, USA
Gillian Lyons-Levy
Daniel L Weeks
Rebecca S Hartley
Resource Type: Journal article
Publication Details: Cell cycle (Georgetown, Tex.), Vol.4(10), pp.1396-1402
DOI: 10.4161/cc.4.10.2035
PMID: 16131839
NLM abbreviation: Cell Cycle
ISSN: 1538-4101
eISSN: 1551-4005
Publisher: United States
Grant note: R01 CA095898 / NCI NIH HHS R01 GM069944-03 / NIGMS NIH HHS R01 GM069944-01 / NIGMS NIH HHS R55 GM056277-01 / NIGMS NIH HHS R01 GM069944 / NIGMS NIH HHS R01 CA095898-01A1 / NCI NIH HHS R01 GM069944-04 / NIGMS NIH HHS R01 GM069944-05A2 / NIGMS NIH HHS R01 GM069944-02 / NIGMS NIH HHS
Language: English
Date published: 10/2005
Academic Unit: Stead Family Department of Pediatrics; Biochemistry and Molecular Biology
Record Identifier: 9984024514702771

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