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Title: Resistance to acetohydroxamate acquired by slow adaptive increases in urease in cultured tobacco cells

Abstract

Urease activity of tobacco XD cells (1U cells) had undergone a 4-fold increase (4U cells) during a year of growth on urea. A clone of 4U cells gave rise to 12U cells during another year of growth on urea. The doubling time of 12U cells on urea is 2.2 days, compared to about 4 days for 1U cells, while 1U and 12U cells double in 2 days on nitrate. Acetohydroxamic acid (AHA), a specific inhibitor/reversible inactivator of jack bean urease, affects tobacco cells urease similarly. Fifty per cent inhibition of growth by AHA occurred at 20 micromolar in 1U cells growing on urea and at 165 micromolar in 12U cells growing on urea, but at 600 micromolar for either 1U or 12U cells growing on nitrate. When 12U cells were grown on urea with 100 micromolar AHA, extractable urease activity decreased 80% within 2.5 hours and remained at this level for 2 weeks; the doubling time increased to 3.7 days, and intracellular urea rose 2-fold, compared to 12U cells grown on urea without AHA. Urease of 12U cells inactivated by AHA in vivo could be reactivated to its pre-AHA level by incubation at 30 C after extraction and separation frommore » free AHA. AHA inhibited incorporation of /sup 15/N from (/sup 15/N) urea into Kjeldahl nitrogen in the cells, in spite of the increased intracellular urea. These results indicate that AHA acts primarily by inhibiting urease action, rather than by inhibition of formation of urease protein or of uptake of urea. Because 12U cells are 8 times more tolerant of AHA than 1U cells, it is likely that growth on urea in the presence of AHA should select strongly for cells with high urease.« less

Authors:
;
Publication Date:
Research Org.:
Michigan State Univ., East Lansing
OSTI Identifier:
6461405
DOE Contract Number:  
AC02-76ER01338
Resource Type:
Journal Article
Journal Name:
Plant Physiol.; (United States)
Additional Journal Information:
Journal Volume: 67:6
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; CARBONIC ACID ESTERS; BIOLOGICAL EFFECTS; UREASE; ENZYME ACTIVITY; BIOCHEMISTRY; CELL CULTURES; CLONING; COMPARATIVE EVALUATIONS; DNA; ENZYME INHIBITORS; GENES; GRAPHS; GROWTH; HYDROLYSIS; METABOLISM; NICOTIANA; NITROGEN 15; PLANT CELLS; TRACER TECHNIQUES; UREA; AMIDES; AMIDINASES; CARBONIC ACID DERIVATIVES; CHEMICAL REACTIONS; CHEMISTRY; DECOMPOSITION; ENZYMES; ESTERS; HYDROLASES; ISOTOPE APPLICATIONS; ISOTOPES; LIGHT NUCLEI; LYSIS; NITROGEN ISOTOPES; NON-PEPTIDE C-N HYDROLASES; NUCLEI; NUCLEIC ACIDS; ODD-EVEN NUCLEI; ORGANIC COMPOUNDS; ORGANIC NITROGEN COMPOUNDS; PLANTS; SOLVOLYSIS; STABLE ISOTOPES; 550201* - Biochemistry- Tracer Techniques; 550301 - Cytology- Tracer Techniques

Citation Formats

Yamaya, T, and Filner, P. Resistance to acetohydroxamate acquired by slow adaptive increases in urease in cultured tobacco cells. United States: N. p., 1981. Web. doi:10.1104/pp.67.6.1133.
Yamaya, T, & Filner, P. Resistance to acetohydroxamate acquired by slow adaptive increases in urease in cultured tobacco cells. United States. https://doi.org/10.1104/pp.67.6.1133
Yamaya, T, and Filner, P. 1981. "Resistance to acetohydroxamate acquired by slow adaptive increases in urease in cultured tobacco cells". United States. https://doi.org/10.1104/pp.67.6.1133.
@article{osti_6461405,
title = {Resistance to acetohydroxamate acquired by slow adaptive increases in urease in cultured tobacco cells},
author = {Yamaya, T and Filner, P},
abstractNote = {Urease activity of tobacco XD cells (1U cells) had undergone a 4-fold increase (4U cells) during a year of growth on urea. A clone of 4U cells gave rise to 12U cells during another year of growth on urea. The doubling time of 12U cells on urea is 2.2 days, compared to about 4 days for 1U cells, while 1U and 12U cells double in 2 days on nitrate. Acetohydroxamic acid (AHA), a specific inhibitor/reversible inactivator of jack bean urease, affects tobacco cells urease similarly. Fifty per cent inhibition of growth by AHA occurred at 20 micromolar in 1U cells growing on urea and at 165 micromolar in 12U cells growing on urea, but at 600 micromolar for either 1U or 12U cells growing on nitrate. When 12U cells were grown on urea with 100 micromolar AHA, extractable urease activity decreased 80% within 2.5 hours and remained at this level for 2 weeks; the doubling time increased to 3.7 days, and intracellular urea rose 2-fold, compared to 12U cells grown on urea without AHA. Urease of 12U cells inactivated by AHA in vivo could be reactivated to its pre-AHA level by incubation at 30 C after extraction and separation from free AHA. AHA inhibited incorporation of /sup 15/N from (/sup 15/N) urea into Kjeldahl nitrogen in the cells, in spite of the increased intracellular urea. These results indicate that AHA acts primarily by inhibiting urease action, rather than by inhibition of formation of urease protein or of uptake of urea. Because 12U cells are 8 times more tolerant of AHA than 1U cells, it is likely that growth on urea in the presence of AHA should select strongly for cells with high urease.},
doi = {10.1104/pp.67.6.1133},
url = {https://www.osti.gov/biblio/6461405}, journal = {Plant Physiol.; (United States)},
number = ,
volume = 67:6,
place = {United States},
year = {1981},
month = {6}
}