In vivo repopulation of cytoplasmically gene transferred hematopoietic cells by temperature-sensitive mutant of recombinant Sendai viral vector

2007 ◽  
Vol 361 (3) ◽  
pp. 811-816 ◽  
Author(s):  
Kumi Yoshida ◽  
Yoshikazu Yonemitsu ◽  
Sakura Tanaka ◽  
Shuro Yoshida ◽  
Satoko Shibata ◽  
...  
Keyword(s):  
Viral Vector ◽  
Hematopoietic Cells ◽  
Temperature Sensitive ◽  
Sensitive Mutant ◽  
Temperature Sensitive Mutant

Directionality of microtubule assembly: an in vivo study with the ciliate Tetrahymena

1978 ◽  
Vol 33 (1) ◽  
pp. 227-234
Author(s):  
S.F. Ng
Keyword(s):  
Tetrahymena Thermophila ◽  
Recessive Gene ◽  
Microtubule Assembly ◽  
In Vivo Study ◽  
Temperature Sensitive ◽  
Sensitive Mutant ◽  
Posterior Portion ◽  
Temperature Sensitive Mutant

A temperature-sensitive mutant homozygous for the recessive gene molb in Tetrahymena thermophila offers opportunity for studying the direction of microtubule assembly in vivo. At 39 degrees C the mutant fails to divide properly; the 2 daughter animals remain attached and bend over each other. As revealed by protargol staining, the bending results in acute turning and breaking of some of the longitudinal microtubular bands close and parallel to the surface. Hence, 2 broken microtubular ends are available for study of the problem of directionality of microtubule assembly, by assessing which of the 2 ends regenerates. In most cases the posterior portion of the longitudinal microtubular band regenerates. The present study hence supports the conclusion based on in vitro observation in other systems that microtubule assembly is predominantly unidirectional.

scholarly journals Kei1: A Novel Subunit of Inositolphosphorylceramide Synthase, Essential for Its Enzyme Activity and Golgi Localization

2009 ◽  
Vol 20 (20) ◽  
pp. 4444-4457 ◽  
Author(s):  
Keisuke Sato ◽  
Yoichi Noda ◽  
Koji Yoda
Keyword(s):  
Inositol Phosphate ◽  
Specific Inhibitor ◽  
Growth Defect ◽  
Temperature Sensitive ◽  
Sensitive Mutant ◽  
Temperature Sensitive Mutant ◽  
Golgi Localization ◽  
Head Groups ◽  
Aureobasidin A

Fungal sphingolipids have inositol-phosphate head groups, which are essential for the viability of cells. These head groups are added by inositol phosphorylceramide (IPC) synthase, and AUR1 has been thought to encode this enzyme. Here, we show that an essential protein encoded by KEI1 is a novel subunit of IPC synthase of Saccharomyces cerevisiae. We find that Kei1 is localized in the medial-Golgi and that Kei1 is cleaved by Kex2, a late Golgi processing endopeptidase; therefore, it recycles between the medial- and late Golgi compartments. The growth defect of kei1-1, a temperature-sensitive mutant, is effectively suppressed by the overexpression of AUR1, and Aur1 and Kei1 proteins form a complex in vivo. The kei1-1 mutant is hypersensitive to aureobasidin A, a specific inhibitor of IPC synthesis, and the IPC synthase activity in the mutant membranes is thermolabile. A part of Aur1 is missorted to the vacuole in kei1-1 cells. We show that the amino acid substitution in kei1-1 causes release of Kei1 during immunoprecipitation of Aur1 and that Aur1 without Kei1 has hardly detectable IPC synthase activity. From these results, we conclude that Kei1 is essential for both the activity and the Golgi localization of IPC synthase.

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