~IoNITOR
Identification of the cystic fibrosis
gene: chromosome walking and
jumping
J.M. ROMMENS ET AL.
Science 245, 1059-1065
Identification of the cystic fibrosis
gene: cloning and characterization of
complementary DNA
J.R. RIORDAN ET AL.
Science 245, 1066-1073
Identification of the cystic fibrosis
gene: genetic analysis
B-S. KEREM ET AL.
Science 245, 1073-1080
The cystic fibrosis (CF) gene has
finally b e e n cloned! A large collabo-
rative effort has identified the prob-
able locus, characterized its gene
MyoD expression in the forming
somites is an early response to meso-
derm induction in Xenopus embryos
N.D. HOPWOOD~ A. PLUCK AND
J.B. GURDON
EMBOJ 8, 3409-3417
MleoD was first identified as a cDNA
that could stably convert mouse
fibroblasts in tissue culture into
myoblasts. It appears to encode a se-
quence-specific DNA-binding protein
related to the c-myc gene product.
H o p w o o d et al. have n o w isolated
the Xenopus h o m o l o g u e of MyoD,
and analysed its expression during
frog embryogenesis. During gastrula-
tion, extensive dorsal m e s o d e r m cell
m o v e m e n t s lead to the formation of
The multivulva phenotype of certain
C. elegans mutants results from defects
in two functionally redundant
pathways
E.L. FERGUSON AND H.R. HORVITZ
Genetics 123, 109-121
The vulva of C. elegans forms from
three of six precursor cells, in
response to an inductive signal from
the anchor cell of the somatic gonad.
At least 11 genes are k n o w n to affect
the fates of the vulval precursor cells.
For example in Muv (multivulva)
mutants all six cells express vulval as
apposed to non-vulval fates, giving
rise to several vulva-like protrusions.
The CB1322 C. elegans strain is a
Muv mutant, but its defects d e p e n d
on the presence of two mutations;
either mutation on its o w n does not
give rise to a Muv phenotype.
Ferguson and Horvitz have n o w
identified eight n e w genes that also
give rise to silent Muv mutations;
they do not give a Muv phenotype
©1989 Elsevier Science Publishers Ltd (UK) 01(~8 - 9479.89 $02.00
product and determined h o w the
product differs in CF patients.
Linkage analysis had previously
localized CF to chromosome band
7q31. From two closely linked flank-
ing markers, the group applied a bat-
tery of techniques to clone the CF
locus, in particular carrying out chro-
mosome jumping and walking over
280 kb. Within this region a search
was begun to identify potential gene
sequences by a variety of techniques,
including the detection of regions
conserved among different species.
This latter method pinpointed four
interesting regions, one of which led
eventually to the isolation of cDNA
clones spanning 61 kb and potentially
encoding a 1480 amino acid protein.
The similarity between the predicted
somites either side of the dorsal mid-
line; MyoD is expressed in the future
somites of the gastrula m e s o d e r m -
much earlier than previously identi-
fied muscle marker genes (e.g. the
actin genes). Surprisingly, MyoD is
not expressed in embryonic heart
muscle cells, even though they
express skeletal actin isoforms, indi-
cating that striated muscle genes, in
some cells at least, have an alterna-
tive activation mechanism. The beauty
of this system lies in the marriage of
a mouse culture system in which
interesting molecules can be readily
identified, and an 'in vivo' system
(the Xenopus embryos) in which the
developmental effects of the mol-
ecules can be viewed. /a
on their o w n but do in combination
with one of the CB1322 mutations.
Interestingly, these mutations fall into
two classes - a combination of two
mutations from within a class remain
silent, but a mutation from one class
combined with one from the other
class leads to a Muv worm. While
genetic redundancy has been seen at
the level of gene families (e.g. either
of two yeast ras genes is effective),
this is the first evidence that redun-
dancy can also be apparent at the
level of gene pathways. General
genetic screens are unlikely to pick
up genes in either of two redundant
pathways, as the other pathway will
take over and produce a 'wild-type'
structure. Some of the silent Muv
genes studied by Ferguson and
Horvitz may have functions b e y o n d
determination of vulval cell fates. A
molecular analysis should determine
what role the pathways play in vulval
determination and why they are
redundant.
TIG NOVEMBER1989 VOL. 5 NO. 11
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m
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protein, CFTR (CF transmembrane
conductance regulator), and the multi-
drug resistance P-glycoprotein and
other membrane-associated proteins
suggests that it is involved in the
movement of ions across membranes.
This is consistent with known defects
in anion transport in CF patients.
Interestingly, 70% of all CF
patients share the same mutation -
the deletion of a phenylalanine
residue at position 508; many other
different mutations form the remain-
der. The identification of the CFlocus
should benefit those searching for
treatment for the disorder; in ad-
dition, the strategies used in the
search should find application in
other genetic diseases for which no
protein product has been identified.
Identification of a mammalian protein
that binds specificallyto DNA
containing methylated CpGs
R.R. MI~I~HANE T A I ,
Cell 58, 499-507
DNA methylation in mammalian cells
is associated with transcriptional
inactivation. Only CpG dinucleotides
are ever modified in this way (on the
cytosine residue), and up to 90% may
be methylated. Many of the non-
methylated CpGs are found in clus-
ters (CpG islands) close to genes,
while the remainder tend to be dis-
persed throughout the genome,
along with the methylated dinu-
cleotides. The inhibition of transcrip-
tion by DNA methylation implies
either that transcription factors are
unable to bind methylated DNA or
that some other factors preferentially
interact with methylated DNA and
consequently hinder transcription of
DNA in that region. At least one tran-
scription factor (Spl) can bind to
either methylated or demethylated
promoters and stimulate transcrip-
tion, favouring the second possibility.
Now Meehan et al. have isolated a
protein (methyl-CpG-binding protein,
MeCP) that can bind to numerous
DNA sequences as long as they con-
tain multiple methylated CpGs. It is
not yet certain that MeCP binds to
dispersed CpGs in the cell, although
competition experiments and the fact
that methylated CpGs are more resis-
tant to nuclease digestion in the cell
(reported in an accompanying paper;
F. Antequera et al. Cell 58, 509-517)
suggest that it does. It is also not
clear h o w binding of MeCP might act
(perhaps by direct interference with
transcription factors or by induction
of an inactive chromatin configura-
tion).