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. 2023 Aug 22:2023:10.17912/micropub.biology.000936.
doi: 10.17912/micropub.biology.000936. eCollection 2023.

clifford B.4.1 , an allele of CG1603 , causes tissue overgrowth in the Drosophila melanogaster eye

Reagan R Nowaskie  1 , Ashley Kitch  1 , Abby Adams  2 , Abinaya Anandaraj  3 , Ethan Apawan  4 , Liliana Bañuelos  2 , Cassandra J Betz  3 , Julia M Bogunia  2 , Nicholas Buechlein  1 , Morgan R Burns  2 , Hayley A Collier  2 , Zach Collins  2 , Kynzie Combs  1 , Vana D Dakarian  2 , Abigail Daniel  1 , Conrad M De Jesus Iii  1 , John D Erickson  1 , Bianca Estrada  2 , Kevin Estrada  2 , Sydney Fields  2 , Maya Gabriel  4 , Rosario M Garcia  2 , Sylvia Gitamo  3 , Emma Granath  2 , Sabrina N Hardin  4 , Emily Hattling  3 , Alexandra Vl Henriquez  2 , Destiny Hernandez  4 , Luke Johnson  3 , Annie H Kim  1 , Lillian K Kolley  1 , Katelynn M Larue  2 , Erin Lockwood  2 , Nelia Longoria  4 , Cassandra Lopez  2 , Rosario C Lopez-Roca Fernandez  4 , Sofia Lozano  4 , Carissa Manthie  3 , Trinity May  3 , Zorah Mehrzad  1 , Itzel Mendoza  2 , Somya Mohan  4 , Claylan Mounthachak  3 , Merveille Muyizere  2 , Margaret R Myers  2 , Jayce Newton  2 , Amarachi Nwawueze  2 , Ariana J Paredes  1 , Marissa N Pezdek  2 , Hoang Phat Nguyen  3 , Nadia Pobuda  3 , Sahar Sadat  2 , Johnathon J Sailor  2 , David Santiago  2 , Madison Sbarbaro  2 , David E Schultz Iii  1 , Anahita N Senobari  2 , Emma M Shouse  1 , Sarah M Snarski  2 , Estefanie Solano  2 , Naomi Solis Campos  2 , Elnora Stewart  1 , Jessica Szczepaniak  2 , Michael Tejeda  2 , Dominic F Teoli  2 , Michael Tran  4 , Nishita Trivedi  1 , Laurita Uribe Aristizabal  2 , Bryan Z Vargas  2 , Kenneth W Walker Iii  4 , Joseph Wasiqi  2 , Joyi Wong  4 , Adira Zachrel  2 , Hemin P Shah  2 , Elizabeth Small  2 , Charlie T Watts  1 , Paula Croonquist  3 , Olivier Devergne  2 , Amy K Jones  4 , Elizabeth E Taylor  2 , Jacob D Kagey  5 , Julie A Merkle  1
Affiliations

clifford B.4.1 , an allele of CG1603 , causes tissue overgrowth in the Drosophila melanogaster eye

Reagan R Nowaskie et al. MicroPubl Biol. .

Abstract

Mutant B.4.1 , generated via EMS mutagenesis in Drosophila melanogaster , was studied by undergraduate students participating in the Fly-CURE. After inducing genetically mosaic tissue in the adult eye, B.4.1 mutant tissue displays a robust increase in cell division and a rough appearance. Complementation mapping and sequence analysis identified a nonsense mutation in the gene CG1603 , which we named clifford ( cliff ) due to observed increases in red-pigmented mutant tissue compared to controls. cliff encodes a zinc finger-containing protein implicated in transcriptional control. RNAi knockdown of cliff similarly results in rough eyes, confirming a role for Cliff in eye development.

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Conflict of interest statement

The authors declare that there are no conflicts of interest present.

Figures

Figure 1. <b> Characterization of the <i> cliff <sup>B.4.1</sup> </i> mutation by phenotypic analysis, complementation mapping, and genetic sequencing </b>
Figure 1. Characterization of the cliff B.4.1 mutation by phenotypic analysis, complementation mapping, and genetic sequencing
(A-B) The FLP/FRT genetic system was used to induce mitotic recombination in the adult Drosophila eye to examine the B.4.1 mutant phenotype compared to controls. After mitotic recombination, control eyes ( A , genotype ey>Flp , w – ; FRT42D, Dark 82 /FRT42D ) exhibited a higher percentage of wildtype non-pigmented tissue (arrowhead) compared to eyes of B.4.1 mutant flies ( B , arrowhead; genotype ey>Flp , w – ; FRT42D , cliff B.4.1 , Dark 82 / FRT42D ), which displayed nearly entirely pigmented mutant tissue ( B, arrow). (C-E) RNAi knockdown of CG1603 with GMR-GAL4 results in rough eyes, phenocopying cliff B.4.1 mutant clones in the eye. (C) GMR-GAL4 control eye. (D-E) Two independent RNAi lines targeting CG1603 ( cliff ) result in rough eyes under the control of GMR-GAL4 . (F) Map of chromosome 2R showing the deficiency lines (red bars) that failed to complement the B.4.1 mutation and yielded a region of overlap at chromosomal location 2R:7493197..7533553. Image adapted from JBrowse on FlyBase, release FB2023_03 (Gramates et al. 2022). (G-H) The nucleotide locus of the B.4.1 mutation was identified by Sanger sequence analysis. (G) The control sequence (genotype w – ; FRT42D, Dark 82 /CyO ) contained a single G peak at position 2R:7,516,167, which is within the coding region of CG1603 . (H) The mutant sequence (genotype w – ; FRT42D , cliff B.4.1 , Dark 82 /CyO ) contained a heterozygous peak revealing a nucleotide change from G to A at this position (*). (I) Alignment of wildtype (WT) and B.4.1 mutant Clifford (Cliff) protein. The nonsense mutation (*) identified in the B.4.1 mutant line at amino acid 32 (W, tryptophan in wildtype) is indicated (blue box).
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