O. Staub et al., IMMUNOLOCALIZATION OF THE UBIQUITIN-PROTEIN LIGASE NEDD4 IN TISSUES EXPRESSING THE EPITHELIAL NA+ CHANNEL (ENAC), American journal of physiology. Cell physiology, 41(6), 1997, pp. 1871-1880
The epithelial Na+ channel (ENaC) was previously shown to be expressed
in several Na+- and fluid-absorbing epithelia, particularly those of
the kidney, colon, and lung. We have recently identified the ubiquitin
-protein ligase Nedd4 as an interacting protein with ENaC and demonstr
ated that Nedd4 binds by its WW domains to the proline-rich PY motifs
of ENaC. These PY motifs were recently shown to be deleted/mutated in
patients afflicted with Liddle's syndrome, a hereditary form of system
ic renal hypertension. Such mutations cause elevated channel activity
by an increase in channel number/stability at the plasma membrane and
by increased channel opening. We then proposed that Nedd4, by regulati
ng channel stability/degradation, may be a suppressor of ENaC. To test
whether Nedd4 is localized to those tissues/regions that express ENaC
, we performed immunocytochemical analysis of rat Nedd4 (rNedd4) distr
ibution in rat kidney, colon, and lung tissues. Our results show that,
in the kidney, rNedd4 is primarily localized to the cortical collecti
ng tubules and outer and inner medullary collecting ducts. These tubul
ar segments were previously shown to express ENaC. The epithelium lini
ng medullary calyxes was also intensely stained, and microvillar borde
rs of proximal convoluted tubules expressed variable amounts of rNedd4
. In the lung, rNedd4 was mainly expressed in the epithelia lining the
airways, in the submucosal glands and ducts, and in the distal respir
atory epithelium. These sites resemble the pattern of ENaC expression.
In contrast, in the distal colon, rNedd4 was strongly expressed in th
e epithelia lining the crypts but not in the ENaC-expressing surface e
pithelium. Low-salt diet (to elevate serum aldosterone levels) had no
effect on rNedd4 distribution in the kidney or colon. Thus Nedd4 is co
expressed and likely colocalizes with ENaC in specific regions within
the kidney and lung but not in the colon. We speculate this difference
in colocalization may reflect differences in the regulation of channe
l stability in those tissues.