title: Mice develop normally in the absence of Smad4 nucleocytoplasmic shuttling.
abstract: Smad4 in partnership with R-Smads (receptor-regulated Smads) activates TGF-beta (transforming growth factor-beta)-dependent signalling pathways essential for early mouse development. Smad4 null embryos die shortly after implantation due to severe defects in cell proliferation and visceral endoderm differentiation. In the basal state, Smad4 undergoes continuous shuttling between the cytoplasm and the nucleus due to the combined activities of an N-terminal NLS (nuclear localization signal) and an NES (nuclear export signal) located in its linker region. Cell culture experiments suggest that Smad4 nucleocytoplasmic shuttling plays an important role in TGF-beta signalling. In the present study we have investigated the role of Smad4 shuttling in vivo using gene targeting to engineer two independent mutations designed to eliminate Smad4 nuclear export. As predicted this results in increased levels of Smad4 in the nucleus of homozygous ES cells (embryonic stem cells) and primary keratinocytes, in the presence or absence of ligand. Neither mutation affects Smad4 expression levels nor its ability to mediate transcriptional activation in homozygous cell lines. Remarkably mouse mutants lacking the Smad4 NES develop normally. Smad4 NES mutants carrying one copy of a Smad4 null allele also fail to display developmental defects. The present study clearly demonstrates that Smad4 nucleocytoplasmic shuttling is not required for embryonic development or tissue homoeostasis in normal, healthy adult mice.
authors.affiliation: The Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, UK.
authors.forename: Christine A
authors.initials: CA
authors.lastname: Biondi
authors.forename: Debipriya
authors.initials: D
authors.lastname: Das
authors.forename: Michael
authors.initials: M
authors.lastname: Howell
authors.forename: Ayesha
authors.initials: A
authors.lastname: Islam
authors.forename: Elizabeth K
authors.initials: EK
authors.lastname: Bikoff
authors.forename: Caroline S
authors.initials: CS
authors.lastname: Hill
authors.forename: Elizabeth J
authors.initials: EJ
authors.lastname: Robertson
mesh_terms: D000483:Alleles; D000818:Animals; D001483:Base Sequence; D002467:Cell Nucleus / Q000378:metabolism*; D002478:Cells, Cultured; D003593:Cytoplasm / Q000378:metabolism*; D017931:DNA Primers; D018390:Gene Targeting; D051379:Mice; D018345:Mice, Knockout; D008817:Mice, Mutant Strains; D012326:RNA Splicing; D015398:Signal Transduction; D051901:Smad4 Protein / Q000235:genetics / Q000378:metabolism*; D015533:Transcriptional Activation; D016212:Transforming Growth Factor beta / Q000502:physiology
chemical_list: D017931:DNA Primers; D051901:Smad4 Protein; D016212:Transforming Growth Factor beta