A distinct H2A.X isoform is enriched in Xenopus laevis eggs and early embryos and is phosphorylated in the absence of a checkpoint

David Shechter, Raghu K. Chitta, Andrew Xiao, Jeffrey Shabanowitz, Donald F. Hunt, C. David Allis

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Histone H2A.X is an H2A variant present in multicellular organisms that is specifically phosphorylated on the serine in the C-terminal consensus sequence, canonically "SQEY," in response to DNA damage. We have recently shown the significance of phosphorylation of the penultimate tyrosine for maintenance and processing of the DNA damage response in mammalian cells. Here, we report the identification of distinct H2A.X variants in the eggs and early embryos of the frog Xenopus laevis that contain a C-terminal SQEF, among other changes; we have denoted these proteins as "H2A.XF." H2A.X-F is present only in late-staged oocytes, eggs, and premidblastula transition embryos and is not present in somatic cells. Similar unannotated isoforms were identified in other rapidly developing aquatic species, such as Xenopus tropicalis, goldfish, and zebrafish, and in Arabidopsis and chickpea. Furthermore, we demonstrate by mass spectrometry and phospho-specific antibodies that H2A.X-F is phosphorylated in the absence of exogenous DNA damage, in both actively dividing, unperturbed embryos and cell-free egg extract in the absence and presence of DNA damage and S-phase checkpoint conditions. We propose that this isoform may be involved in modulating the cellular response to the rapid early cell cycles in externally developing species.

Original languageEnglish (US)
Pages (from-to)749-754
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume106
Issue number3
DOIs
StatePublished - Jan 20 2009
Externally publishedYes

Fingerprint

Xenopus laevis
Eggs
DNA Damage
Protein Isoforms
Embryonic Structures
S Phase Cell Cycle Checkpoints
Phospho-Specific Antibodies
Cicer
Goldfish
Consensus Sequence
Zebrafish
Xenopus
Arabidopsis
Anura
Histones
Serine
Oocytes
Ovum
Tyrosine
Mass Spectrometry

Keywords

  • Chromatin
  • DNA damage
  • Histone
  • Variant

ASJC Scopus subject areas

  • General

Cite this

A distinct H2A.X isoform is enriched in Xenopus laevis eggs and early embryos and is phosphorylated in the absence of a checkpoint. / Shechter, David; Chitta, Raghu K.; Xiao, Andrew; Shabanowitz, Jeffrey; Hunt, Donald F.; Allis, C. David.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 106, No. 3, 20.01.2009, p. 749-754.

Research output: Contribution to journalArticle

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AU - Shabanowitz, Jeffrey

AU - Hunt, Donald F.

AU - Allis, C. David

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