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In vivo mapping of the functional regions of the DEAD-box helicase Vasa.

Dehghani M, Lasko P - Biol Open (2015)

Bottom Line: We identified novel domains in the N- and C-terminal regions of the protein that are essential for localization, transposon repression, posterior patterning, and pole cell specification.One such functional region, the most C-terminal seven amino acids, is specific to Vas orthologues and is thus critical to distinguishing Vas from other closely related DEAD-box helicases.Surprisingly, we also found that many eGFP-Vas proteins carrying mutations that would be expected to abrogate DEAD-box helicase function localized to the nuage and posterior pole, and retained the capacity to support oogenesis, although they did not function in embryonic patterning, pole cell specification, grk activation, or transposon repression.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, McGill University, 3649 Promenade Sir William Osler, Montréal, QC H3G 0B1, Canada.

No MeSH data available.


The ability of various egfp-vas constructs to restore abdominal segmentation in vas1 embryos.(A) Hatching rates: the red bars indicate the percentage of embryos hatched after 48 hours, error bars indicate SEM from at least five plates. Between 500-1000 embryos in total were scored for each genotype. (B) ftz expression in vas1 embryos containing various egfp-vas constructs: The y-axis indicates the average number of ftz stripes for each genotype. Error bars show SEM calculated from more than 50 embryos examined for each genotype. In both A and B asterisks show a significant increase or decrease from vas+ (p<0.05).
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f08: The ability of various egfp-vas constructs to restore abdominal segmentation in vas1 embryos.(A) Hatching rates: the red bars indicate the percentage of embryos hatched after 48 hours, error bars indicate SEM from at least five plates. Between 500-1000 embryos in total were scored for each genotype. (B) ftz expression in vas1 embryos containing various egfp-vas constructs: The y-axis indicates the average number of ftz stripes for each genotype. Error bars show SEM calculated from more than 50 embryos examined for each genotype. In both A and B asterisks show a significant increase or decrease from vas+ (p<0.05).

Mentions: vas1 embryos fail to hatch because they lack posterior segments and thus die. We tested the ability of the various eGFP-Vas constructs to rescue embryonic viability to vas1 embryos. Proteins with N-terminal deletions within the interval 15-127 differ little from wild-type eGFP-Vas in their ability to restore viability to vas1 embryos (Fig. 8A). However, embryonic viability decreased about 37% by removing amino acids 3-139 (p = 0.044) and was almost completely abolished by a more extensive N-terminal deletion (Δ3-200). We also found that all seven mutations in the conserved core motifs abrogated the function of Vas with regard to embryonic viability.


In vivo mapping of the functional regions of the DEAD-box helicase Vasa.

Dehghani M, Lasko P - Biol Open (2015)

The ability of various egfp-vas constructs to restore abdominal segmentation in vas1 embryos.(A) Hatching rates: the red bars indicate the percentage of embryos hatched after 48 hours, error bars indicate SEM from at least five plates. Between 500-1000 embryos in total were scored for each genotype. (B) ftz expression in vas1 embryos containing various egfp-vas constructs: The y-axis indicates the average number of ftz stripes for each genotype. Error bars show SEM calculated from more than 50 embryos examined for each genotype. In both A and B asterisks show a significant increase or decrease from vas+ (p<0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4400588&req=5

f08: The ability of various egfp-vas constructs to restore abdominal segmentation in vas1 embryos.(A) Hatching rates: the red bars indicate the percentage of embryos hatched after 48 hours, error bars indicate SEM from at least five plates. Between 500-1000 embryos in total were scored for each genotype. (B) ftz expression in vas1 embryos containing various egfp-vas constructs: The y-axis indicates the average number of ftz stripes for each genotype. Error bars show SEM calculated from more than 50 embryos examined for each genotype. In both A and B asterisks show a significant increase or decrease from vas+ (p<0.05).
Mentions: vas1 embryos fail to hatch because they lack posterior segments and thus die. We tested the ability of the various eGFP-Vas constructs to rescue embryonic viability to vas1 embryos. Proteins with N-terminal deletions within the interval 15-127 differ little from wild-type eGFP-Vas in their ability to restore viability to vas1 embryos (Fig. 8A). However, embryonic viability decreased about 37% by removing amino acids 3-139 (p = 0.044) and was almost completely abolished by a more extensive N-terminal deletion (Δ3-200). We also found that all seven mutations in the conserved core motifs abrogated the function of Vas with regard to embryonic viability.

Bottom Line: We identified novel domains in the N- and C-terminal regions of the protein that are essential for localization, transposon repression, posterior patterning, and pole cell specification.One such functional region, the most C-terminal seven amino acids, is specific to Vas orthologues and is thus critical to distinguishing Vas from other closely related DEAD-box helicases.Surprisingly, we also found that many eGFP-Vas proteins carrying mutations that would be expected to abrogate DEAD-box helicase function localized to the nuage and posterior pole, and retained the capacity to support oogenesis, although they did not function in embryonic patterning, pole cell specification, grk activation, or transposon repression.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, McGill University, 3649 Promenade Sir William Osler, Montréal, QC H3G 0B1, Canada.

No MeSH data available.