Abstract
Recently, human asparagine synthetase has been found to be associated with the mitotic spindle. However, this event cannot be seen in yeast because yeast takes a different cell division process via closed mitosis (there is no nuclear envelope breakdown to allow the association between any cytosolic enzyme and mitotic spindle). To find out if yeast asparagine synthetase can also (but hiddenly) have this feature, the coding sequences of green fluorescent protein (GFP) and nuclear localization signal (NLS) were introduced downstream ofASN1andASN2, encoding asparagine synthetases Asn1p and Asn2p, respectively, in the yeast genome havingmCherrrycoding sequence downstream ofTUB1encoding alpha-tubulin, a building block of the mitotic spindle. The genomically engineered yeast strains showed co-localization of Asn1p-GFP-NLS (or Asn2p-GFP-NLS) and Tub1p-mCherry in dividing nuclei. In addition, an activity-disrupted mutation was introduced toASN1(orASN2). The yeast mutants still exhibited co-localization between defective asparagine synthetase and mitotic spindle, indicating that the biochemical activity of asparagine synthetase is not required for its association with the mitotic spindle. Furthermore, nocodazole treatment was used to depolymerize the mitotic spindle, resulting in lack of association between the enzyme and the mitotic spindle. Although yeast cell division undergoes closed mitosis, preventing the association of its asparagine synthetase with the mitotic spindle, however, by using yeast constructs with re-localized Asn1/2p have suggested the moonlighting role of asparagine synthetase in cell division of higher eukaryotes.
Funder
The Coordinating Center for Thai Government Science and Technology Scholarship Students (CSTS) – The National Science and Technology Development Agency
The National Research Council of Thailand (NRCT) in association with Office of the Permanent Secretary, Ministry of Higher Education, Science, Research and Innovation
The Development and Promotion of Science and Technology Talents Project
Mahidol University
Publisher
Public Library of Science (PLoS)
Cited by
4 articles.
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