Loss of function of the Drosophila Ninein-related centrosomal protein Bsg25D causes mitotic defects and impairs embryonic development

Author:

Kowanda Michelle1,Bergalet Julie2,Wieczorek Michal1,Brouhard Gary1,Lécuyer Éric234,Lasko Paul1ORCID

Affiliation:

1. Department of Biology, McGill University, Montréal, Québec H3G 0B1, Canada

2. RNA Biology Unit, Institut de recherches cliniques de Montréal (IRCM), Montréal, Québec H2W 1R7, Canada

3. Département de Biochimie, Université de Montréal, Montréal, Québec H3T 1J4, Canada

4. Division of Experimental Medicine, McGill University, Montréal, Québec H3A 1A3, Canada

Abstract

ABSTRACT The centrosome-associated proteins Ninein (Nin) and Ninein-like protein (Nlp) play significant roles in microtubule stability, nucleation and anchoring at the centrosome in mammalian cells. Here, we investigate Blastoderm specific gene 25D (Bsg25D), which encodes the only Drosophila protein that is closely related to Nin and Nlp. In early embryos, we find that Bsg25D mRNA and Bsg25D protein are closely associated with centrosomes and astral microtubules. We show that sequences within the coding region and 3′UTR of Bsg25D mRNAs are important for proper localization of this transcript in oogenesis and embryogenesis. Ectopic expression of eGFP-Bsg25D from an unlocalized mRNA disrupts microtubule polarity in mid-oogenesis and compromises the distribution of the axis polarity determinant Gurken. Using total internal reflection fluorescence microscopy, we show that an N-terminal fragment of Bsg25D can bind microtubules in vitro and can move along them, predominantly toward minus-ends. While flies homozygous for a Bsg25D null mutation are viable and fertile, 70% of embryos lacking maternal and zygotic Bsg25D do not hatch and exhibit chromosome segregation defects, as well as detachment of centrosomes from mitotic spindles. We conclude that Bsg25D is a centrosomal protein that, while dispensable for viability, nevertheless helps ensure the integrity of mitotic divisions in Drosophila.

Funder

Canadian Institutes of Health Research

Natural Sciences and Engineering Research Council of Canada

Fonds de Recherche du Québec - Santé

Publisher

The Company of Biologists

Subject

General Agricultural and Biological Sciences,General Biochemistry, Genetics and Molecular Biology

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