The Nature of the wimp Mutation

wimp is a dominant maternal-effect mutation, allelic to mutations in the second largest (140 kD) subunit of RNA polymerase II, that interacts with a specific subset of early genes, including many segmentation genes such as the pair-rule gene hairy. wimp is a change-of-function (antimorphic) mutation that reduces, but does not eliminate, transcription of these interacting genes, such that one wild-type copy of the interacting gene is no longer sufficient for normal development. 

Embryos heterozygous for an interacting mutation, such as hairy, die when derived from a wimp/+ mother.

The reduction in interacting gene product due to the wimp maternal effect is reflected in both protein levels (shown below for the segmentation protein, Ftz) and at the level of transcription as assayed by in situ hybridization.

Embryos derived from wimp/+ mothers crossed to wildtype fathers develop normally, despite the reduction of transcript levels of all interacting genes. In addition to its conditional maternal effect summarized above, wimp is also a recessive lethal mutation: homozygous wimp embryos die with severe cuticle pattern disruptions, suggesting that wimp may affect processes such as cellularization and gastrulation.

We have looked at the interaction of the wimp maternal effect with a variety of different loci, including mutations in maternally- and zygotically-acting loci, by scoring for embryonic lethality.

Using the wimp Mutation to Identify Maternal Effect Lethals

Based on our survey of interactions of early developmental genes with the wimp maternal effect, we reasoned that genetic screens using wimp would identify mutations in a variety of early developmental processes including fertilization, nuclear cleavage divisions, nuclear migration, pattern formation, sex determination, cellularization and gastrulation. Although wimp exhibits interaction specificity such that it does not interact with all mutations, it renders many interacting recessive mutations pseudodominant and greatly eases the identification of novel maternal loci. Importantly, wimp allows recovery of mutations in genes that are required both maternally and zygotically, a class of mutation that is lost in conventional screens because of recessive lethality. wimp also allows identification of maternally-acting mutations that are required for cell viability (mutations where germline clones cannot be recovered).

With this in mind, we set out to identify wimp-interacting mutations with crucial roles in Drosophila oogenesis or early embryogenesis that define new pathways of regulation. We have screened approximately 3500 lethal P-element lines (lines from Bloomington, Berkeley, and I. Kiss) and have isolated 98 mutations representing 92 complementation groups that interact maternally with wimp and that are essential for early development. We have put these mutations into different classes on the basis of their cuticle phenotypes.

To get a better idea of the kinds of gene products/processes being disrupted by these mutations, we rescued the DNA sequences flanking the P-element for 85 of the mutant lines. Using primers specific to the P-element ends, we obtained roughly one kilobase of sequence flanking each side of the P-element and then searched for sequence homologies. Roughly 30% of the sequences showed homology to known genes or protein motifs.

• Adh distal factor-1
• GATA2 factor
• trx-like (GAGA factor)
• cap 'n collar (bZIP)
• Lighten up (helicase)
• pointed (ETS domain)
• vein (EGF repeats)
• protein phosphatase 2a (55kD subunit)
  

• protein phosphatase 2a (65kD subunit)
• protein phosphatase 2a (cat. subunit)
• FTSJ (cell division protein)
• Sir2 (deacetylase)
• PCNA
• cyclin A
• Rho (GTPase)
• ß-tubulin

• E-cadherin
• unchained
• poly-A binding protein
• trachealess (bHLH-PAS)
• saxophone (TGFß type I receptor)
• expanded (SH3 domains)