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Screening of genes involved in cell migration in Dictyostelium.

Abstract：

:A single cell of wild-type Dictyostelium discoideum forms a visible colony on a plastic dish in several days, but due to enhanced cell migration, amiB-null mutant cells scatter over a large area and do not form noticeable colonies. Here, with an aim to identify genes involved in cell migration, we isolated suppresser mutants of amiB-null mutants that restore the ability to form colonies. From REMI (restriction enzyme-mediated integration)-mutagenized pool of double-mutants, we identified 18 responsible genes from them. These genes can be categorized into several biological processes. One cell line, Sab16 (Suppressor of amiB) was chosen for further analysis, which had a disrupted phospholipase D pldB gene. To confirm the role of pldB gene in cell migration, we knocked out the pldB gene and over-expressed gfp-pldB in wild-type cells. GFP-PLDB localized to plasma membrane and on vesicles, and in migrating cells, at the protruding regions of pseudopodia. Migration speed of vegetative pldB-null cells was reduced to 73% of that of the wild-type. These results suggest that PLDB plays an important role in migration in Dictyostelium cells, and that our screening system is useful for the identification of genes involved in cell migration.

journal_name

Exp Cell Res

journal_title

Experimental cell research

authors

Nagasaki A,Uyeda TQ

doi

10.1016/j.yexcr.2007.12.002

subject

Has Abstract

pub_date

2008-03-10 00:00:00

pages

1136-46

issue

5

eissn

0014-4827

issn

1090-2422

pii

S0014-4827(07)00577-0

journal_volume

314

pub_type

杂志文章

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