The tripartite motif of nuclear factor 7 is required for its association with transcriptional units

Brent Beenders, Peter Lawrence Jones, Michel Bellini

Research output: Contribution to journalArticle

Abstract

In amphibian oocytes, the maternal nuclear factor NF7 associates with the elongating pre-mRNAs present on the numerous lateral loops of the lampbrush chromosomes. Here, we have purified NF7 from an oocyte extract by using a combination of ion-exchange chromatography and gel filtration chromatography and demonstrated for the first time that nucleoplasmic NF7 exists primarily as free homotrimers. We confirmed the in vivo homotrimerization of NF7 by using a glutaraldehyde cross-linking assay, and we further showed that it only requires the coiled-coil domain of the NF7 tripartite motif/RBCC motif. Interestingly, we also obtained evidence that NF7 is recruited to the nucleus as a homotrimer, and expression of several mutated forms of NF7 in oocytes demonstrated that both the coiled coil and B box of NF7 are required for its chromosomal association. Together, these data strongly suggest that the interaction of NF7 with the active transcriptional units of RNA polymerase II is mediated by a trimeric B box. Finally, and in agreement with a role for NF7 in pre-mRNA maturation, we obtained evidence supporting the idea that NF7 associates with Cajal bodies.

Original languageEnglish (US)
Pages (from-to)2615-2624
Number of pages10
JournalMolecular and cellular biology
Volume27
Issue number7
DOIs
StatePublished - Apr 1 2007

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Oocytes
RNA Precursors
RNA Polymerase II
Ion Exchange Chromatography
Glutaral
Amphibians
Gel Chromatography
Chromosomes
Mothers

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

The tripartite motif of nuclear factor 7 is required for its association with transcriptional units. / Beenders, Brent; Jones, Peter Lawrence; Bellini, Michel.

In: Molecular and cellular biology, Vol. 27, No. 7, 01.04.2007, p. 2615-2624.

Research output: Contribution to journalArticle

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