IncreasingtubC ?-tubulin synthesis by placing it under the control of abenA ?-Tubulin upstream sequence causes a reduction inbenA ?-tubulin level but has no effect on microtubule function
✍ Scribed by May, Gregory S. ;Waring, Richard B. ;Morris, N. Ronald
- Publisher
- John Wiley and Sons
- Year
- 1990
- Tongue
- English
- Weight
- 753 KB
- Volume
- 16
- Category
- Article
- ISSN
- 0886-1544
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✦ Synopsis
Abstract
We have constructed a chimeric β‐tubulin gene that places the structural gene for the __tub__C β‐tubulin of Axpergillus nidulans under the control of the __ben__A β‐tubulin promoter. Introduction of cither this chimeric gene or a second wild‐type __ben.__A gene into a benomyl‐resistant __ben__A22 strain causes it to become benomyl sensitive, indicating that the introduced genes are functional. Analysis of the tubulin proteins synthesized in __ben__A22 strains into which a second wild‐type __ben__A β‐tubulin gene was transformed showed that the total amount of β‐tubulin protein was the same as in the parental strain with a single __ben__A gene. Thus the level of β‐tubulin must be regulated. This was also true of transformants carrying an extra copy of the chimeric β‐tubulin gene. The total amount of β‐tubulin was the same as in the parental strain. Two‐dimensional gel analysis showed that the endoge‐nous __ben__A22 and the introduced chimeric __tub__C gene contributed equally to the total β‐tubulin pool. Th; fact that one‐half of the __ben__A β‐tubulin could be replaced by __tub__C β‐tubulin with no effect on the growth of the cells suggests that the __ben__A and __tub__C β‐tubulins are functionally interchangeable.