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and they demonstrate that the regulation of P. M., Ciechanover, A. & Schwartz, A. L. J. Biol. Chem. 1647–1656 (1995).
Genetic studies have conclusively linked connexin channels to human diseases, but the nature of the signals that are disrupted
by channel mutations has remained elusive. A recent study has taken advantage of a deafness-causing mutation to suggest that
permeability to inositol trisphosphate, the Ca2+-mobilizing messenger, is crucial for normal hearing.
The first genetic disease associated with con- Although inherited deafness is genetically where it is partly taken up by cochlear support-
nexin mutations was discovered about ten years heterogeneous, mutations in the gene encod- ing cells5. Hence, Cx26 and Cx30 intercellular
ago, and since then eight more connexin genes ing connexin 26 (Cx26) have been shown to channels between epithelial cells are believed
have been implicated in human hereditary dis- account for a large proportion of cases in every to delineate an intercellular pathway to remove
orders affecting different organs1. Despite all the population tested, and two other connexins, the excess K+ that results from auditory signals,
information provided by animal models and by Cx30 and Cx31, have also been linked to sen- providing a spatial buffering function that is
electrophysiological studies, a notable progress sorineural hearing loss3. In the cochlea, which similar to astrocytic gap junctions in the brain.
into understanding the sequence of events that is the auditory organ, Cx26 and Cx30 proteins Such a role for gap junctions in the cochlea has
occurs in connexin-based disorders has been exhibit overlapping expression patterns4 that been further supported by the finding that mice
lacking. One of the reasons is perhaps inherent define two main communication compart- with a targeted deletion of Cx26 in the epithe-
to the uniqueness of connexin channels, which ments (Fig. 1): an epithelial network, compris- lial network were deaf due to degeneration of
allow the exchange not only of ions, but also of ing supporting cells and adjacent epithelial the organ of Corti6. Because cell death occurred
small molecules between the cytoplasmic envi- cells, and a connective network, both of which at the time of hearing onset, it was thought to
ronment of neighbouring cells. Although the participate in the regulation of the unique ionic result from impaired K+ disposal.
finding that most connexin mutations result in composition of extracellular fluids. There are some problems with this K+
a loss of function suggests that it is a deficit of A key aspect of the physiology of the mam- hypothesis. The foremost being the presence
intercellular communication that leads to the malian cochlea is the ionic composition of in the same epithelial network of another con-
pathological phenotype1, a lingering question the extracellular environment that surrounds nexin, Cx30, which is perfectly capable of car-
has remained unanswered: what is it exactly the hair cells, the cells that sense sound and rying K+ currents7, but cannot functionally
that needs to be exchanged? Ions? Metabolites? initiate signals to afferent nerve fibres. Their compensate for the absence of Cx26 (ref. 6).
Second messengers? On page 63 of this issue2, basolateral surface is bathed in perilymph, a The inability of other connexins to function-
a study by Beltramello et al. has delivered the fluid with normal extracellular salt composi- ally compensate for specific connexin mutants
first answer by pinning down one of the usual tion (high Na+, low K+), but their apical surface is a common feature of connexin-based disor-
suspects, the Ca2+-releasing messenger inosi- is contacted by endolymph, which has a salt ders (the crystalline lens is another example1),
tol trisphosphate (Ins(1,4,5)P3), thus provid- composition that resembles intracellular fluids and has been interpreted as an indication that
ing a plausible hypothesis to account for the (high K+, low Na+). Another unique feature is a specific complement of connexin channels
pathogenesis of DFNB1, the most frequent the very high trans-epithelial potential between is required to meet the physiological needs of
form of deafness. the endolymphatic and perilymphatic compart- a specific tissue. We now know that all con-
ments, which is necessary to trigger the influx nexins are not made equal; there are differ-
Roberto Bruzzone and Martine Cohen-Salmon are in of K+ into hair cells that depolarizes them after ences in size and ionic selectivity, differences
the Department of Neuroscience, Institut Pasteur,
75015 Paris, France. sound stimulation. K+ is, in turn, transported in the rules of compatibility between available
e-mail: bruzzone@pasteur.fr into the interstitial space of the organ of Corti, partners, and distinct gating mechanisms.
K+ ions and, further, to excitotoxic death of hair inactivation of Cx26 in the epithelial network. 1. Gerido, D. A. & White, T. W. Biochim. Biophys. Acta
1662, 159–170 (2004).
and supporting cells. Conversely, inactivation of Cx30, which has 2. Beltramello, M. et al. Nature Cell Biol. 7, 63–69
Of course, there are still a number of unre- a low Ins(1,4,5)P3 permeability and cannot (2005).
3. Petit, C., Levilliers, J. & Hardelin, J. P. Annu. Rev.Genet.
solved issues that should be addressed to vali- functionally compensate for the loss of Cx26, 35, 589–646 (2001).
date this model. First of all it should be borne should not interfere with Ca2+ waves. Finally, 4. Lautermann, J. et al. Cell Tissue Res. 294, 415–420
(1998).
in mind that, although the recorded Ca2+ replacement of Cx26 with V84L should gener- 5. Boettger, T. et al. Nature 416, 874–878 (2002).
6. Cohen-Salmon, M. et al. Curr. Biol. 12, 1106–1111
waves resemble those triggered by mechani- ate mice with hearing loss. (2002).
cal damage or pharmacological stimulation It is tempting to speculate that defects in 7. Valiunas, V. et al. J. Physiol. 519, 631–644 (1999).
8. Bevans, C. G., Kordel, M., Rhee, S. K. & Harris, A. L. J.
of purinergic receptors14, there is no evidence the diffusion of second messengers across gap Biol. Chem. 273, 2808–2816 (1998).
that such intercellular signalling between junctions may offer a mechanistic explana- 9. Goldberg, G. S., Lampe, P. D. & Nicholson, B. J. Nature
Cell Biol. 1, 457–457 (1999).
supporting cells occurs physiologically dur- tion in other connexin-based disorders. In 10. Niessen, H. et al. J. Cell Sci. 113, 1365–1372
(2000).
ing audition. In vivo imaging studies will be fact, a defective transfer of cyclic AMP had 11. Bruzzone, R. et al. FEBS Lett. 533, 79–88 (2003).
needed to address this crucial issue. Then, already been suggested15, albeit on the basis 12. Boitano, S., Dirksen, E. R. & Sanderson, M. J. Science
258, 292–295 (1992).
if the model is correct, one would predict of indirect evidence, as the basis of the X- 13. Elfgang, C. et al. J. Cell Biol. 129, 805–817 (1995).
that a reduction in the propagation of Ca2+ linked form of Charcot-Marie-Tooth, which 14. Gale, J. E., Piazza, V., Ciubotaru, C. D. & Mammano, F.
Curr. Biol. 14, 526–529 (2004).
waves should occur in mice with targeted is caused by mutations in Cx32. 15. Oh, S. et al. Neuron 19, 927–938 (1997).
Injected
The role of Wnt proteins in development runs far and wide. Despite these
Uninjected
many contexts in which to study the Wnt pathway, what lies between Wnt
and specific gene expression is still not clear. Most factors in the canonical
Wnt/β-catenin pathway have already been implicated in cell-fate-specific
gene activation, but lately it is non-canonical Wnt targets that have taken
centre stage. Chen et al. (Nature, DOI:10.1038/nature03126; 2004) now
report for the first time a link between Wnt and PKA-dependent CREB b
Uninjected
signalling during myogenesis.
Using antibodies that recognize the active, phosphorylated form of Injected
CREB on mouse embryonic sections, the authors observed that phos-
pho-CREB and the myogenic transcription factors Pax3, MyoD and
Myf5 are expressed in overlapping patterns. This prompted them to
examine whether CREB-null mutants have defects in muscle develop-
ment. In situ staining revealed that in CREB-null embryos, the levels
of each of these transcription factors is reduced compared with wild-
type. Coincidently, the authors also found that two other CREB-family Interfering with CREB family function in vivo inhibits myogenesis. Control (a) or
members, ATF-1 and CREM, were upregulated enough to at least par- ACREB-expressing (b) adenoviruses were microinjected into E9.75 somites between
tially compensate for the CREB-null phenotype. They confirmed this the forelimb and hindlimb on one side of mouse embryos. Pax3 expression (purple)
by injecting an adenoviral inhibitor (Ad–ACREB) that targets all CREB was analysed by in situ hybridization. Reproduced, with permission, from Nature,
family members: the myogenic defects were now much more severe, thus DOI:10.1038/nature03126 © (2004) Macmillan Magazines Ltd.
reinforcing a role for CREB in myogenesis.
The phenotype seen when CREB was lost was highly reminiscent of cascade — that activates CREB-mediated transcription. They also show
that previously observed for mutants lacking Wnt1 or Wnt3a; thus, Chen that the AC-dependent activation of CREB is physiologically relevant
et al. tested whether CREB might form part of the Wnt signalling path- for Wnt-induced myogenesis. Increasing AC activity (or elevating cAMP
way. Similarly to previous reports, both Wnt proteins induced expression levels) specifically enhanced expression of MyoD and Myf5 induced by
of Pax3, MyoD and Myf5 in a co-culture assay. Importantly, Ad–ACREB Wnt1, whereas blocking AC or PKA activity inhibited both Wnt1- and
infection reduced expression of all three myogenic factors, without Wnt7a-mediated myogenesis, both in cultures and in vivo.
affecting expression of the canonical Wnt/β-catenin pathway targets. Thus, the findings by Chen et al. suggest the existence of a novel Wnt-
Consistent with this, elevated phospho-CREB levels were observed in mediated myogenic pathway involving PKA, AC and CREB. Although
response to expression of both Wnt1 and Wnt3a. it is unclear whether CREB directly activates expression of any of the
So can Wnt regulate CREB-induced transcription directly? Chen et al. myogenic genes examined, it constitutes an important addition to the
see that both Wnt1 and Wnt7a potently induce transcription from the vast array of Wnt effectors. The question now is whether this pathway
cyclic AMP-responsive element (CRE) promoter. By testing a barrage extends its range to other developmental programmes, not least of
of inhibitors and dominant-negative mutants, they conclude that it is which, neurogenesis.
non-canonical Wnt signalling — specifically the adenylyl cyclase (AC) MYRTO RAFTOPOULOU