Worm Breeder's Gazette 11(2): 46

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Sequence Analysis of lin-12(d) Mutations

Iva Greenwald

Figure 1

The accompanying figure shows sequence alterations associated with 
eight lin-12(d) alleles.  All eight lin-12(d) mutations examined are 
missense alterations of the protein coding region and are therefore 
likely to be modifying protein activity or increasing protein 
stability.  Thus, the fact that lin-12(d) mutations behave like 
genetic hypermorphs (Greenwald et al., Cell 34: 435 -444,1983) would 
be consistent with two possibilities: either the lin-12(+) allele 
contributes a basal level of lin-12 activity that can augment lin-12(d)
activity, or the lin-12(d) protein transactivates the lin-12(+) 
protein.  We favor the second possibility, in light of genetic studies 
suggesting that lin-12 interacts with itself (Geraldine Seydoux and I.
G., unpublished observations) and biochemical evidence that Notch can 
form dimers (Kidd et al., Genes & Dev.  3:1113-1129,1989).  Our 
current favorite working hypothesis is that intercellular signals 
promote self-association of lin-12, thereby activating a signal 
transduction mechanism.  This model is plausible in view of what is 
known about biochemically-defined receptors: in many cases, the 
association state of receptors in the membrane controls their activity,
and association state is in turn controlled by ligand-binding (
reviewed in Andersen, Nature 337:12,1989).  The lin-12(d) mutations 
could lead to activation in the absence of signals necessary to 
activate lin12(+) protein.  Consistent with this explanation is the 
observation that if all gonadal cells except for Z1.ppp or Z4.aaa are 
ablated in a lin-12(n137) hermaphrodite, it becomes a VU; in contrast, 
in wild type, it would become an AC, presumably because it lacks a 
necessary signal (Seydoux and Greenwald, Cell 57:1237-1245,1989).
[See Figure 1]
Predicted amino acid changes (residue number: change) deduced from 
sequence analysis of different lin-12(d) alleles are shown on a 
schematic depiction of the lin-12 product (Yochem et al., Nature 335: 
547-550,1988).  EGF = EGF-like motifs; T = 'T + Y' encoded motif; LNR 
= lin12/Notch repeated motif; cdc10 = cdc10/SWI6 motif; C = position 
of a pair of cysteines that is conserved in lin-12, 
h.  Kidd et al.  (1989) propose that these 
cysteines mediate the dimerization of Notch, and it is curious that 
they are conserved in a region of the lin-12 protein that shows no 
overt sequence similarity to Notch.

Figure 1