A novel class of unstable 6-thioguanine-resistant cells from dog and human kidneys

Mitchell Turker, Raymond J. Monnat, Ken Ichiro Fukuchi, Patricia A. Johnston, Charles E. Ogburn, Richard E. Weller, James F. Park, George M. Martin

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Thioguanine-resistant primary clones were grown from single cell suspensions obtained from dog and human kidneys by enzymatic digestion. In medium containing a relatively high concentration (10μg/ ml) of thioguanine, thioguanine-resistant primary clones arose from each source at frequencies ranging from 10-4 to 10-5. A reduction in total hypoxanthine uptake was found in the thioguanine-resistant primary clones which had developed in thioguanine medium, consistent with a reduction in hypoxanthine phosphoribosyltransferase activity. When these thioguanine-resistant primary clones were subsequently grown in the absence of thioguanine and assayed for the thioguanine-resistant phenotype and hypoxanthine phosphoribosyltransferase activity, it was found that most were now thioguanine-sensitive and yielded cell free extracts with substantial amounts of hypoxanthine phosphoribosyltransferase activity. In contrast, thioguanine-resistant human clones grown continuously in the presence of thioguanine yielded cell free extracts with little or no detectable hypoxanthine phosphoribosyltransferase activity. Southern blot analysis demonstrated no structural alterations in the hypoxanthine phosphoribosyltransferase gene in thioguanine-resistant primary human kidney clones. These results suggest that a novel mechanism(s) for thioguanine resistance and the control of hypoxanth phosphoribosyltransferase expression may occur in dog and human kidney cells.

Original languageEnglish (US)
Pages (from-to)211-223
Number of pages13
JournalCell Biology and Toxicology
Volume4
Issue number2
DOIs
StatePublished - Jun 1988
Externally publishedYes

Fingerprint

Thioguanine
Dogs
Kidney
Hypoxanthine Phosphoribosyltransferase
Clone Cells
Cell Extracts
Hypoxanthine
Southern Blotting

Keywords

  • hypoxanthine phosphoribosyltransferase
  • kidney primary clones
  • purine analogue resistance

ASJC Scopus subject areas

  • Toxicology
  • Cell Biology

Cite this

Turker, M., Monnat, R. J., Fukuchi, K. I., Johnston, P. A., Ogburn, C. E., Weller, R. E., ... Martin, G. M. (1988). A novel class of unstable 6-thioguanine-resistant cells from dog and human kidneys. Cell Biology and Toxicology, 4(2), 211-223. https://doi.org/10.1007/BF00119247

A novel class of unstable 6-thioguanine-resistant cells from dog and human kidneys. / Turker, Mitchell; Monnat, Raymond J.; Fukuchi, Ken Ichiro; Johnston, Patricia A.; Ogburn, Charles E.; Weller, Richard E.; Park, James F.; Martin, George M.

In: Cell Biology and Toxicology, Vol. 4, No. 2, 06.1988, p. 211-223.

Research output: Contribution to journalArticle

Turker, M, Monnat, RJ, Fukuchi, KI, Johnston, PA, Ogburn, CE, Weller, RE, Park, JF & Martin, GM 1988, 'A novel class of unstable 6-thioguanine-resistant cells from dog and human kidneys', Cell Biology and Toxicology, vol. 4, no. 2, pp. 211-223. https://doi.org/10.1007/BF00119247
Turker, Mitchell ; Monnat, Raymond J. ; Fukuchi, Ken Ichiro ; Johnston, Patricia A. ; Ogburn, Charles E. ; Weller, Richard E. ; Park, James F. ; Martin, George M. / A novel class of unstable 6-thioguanine-resistant cells from dog and human kidneys. In: Cell Biology and Toxicology. 1988 ; Vol. 4, No. 2. pp. 211-223.
@article{3814720e539e469c93e7263ef250465f,
title = "A novel class of unstable 6-thioguanine-resistant cells from dog and human kidneys",
abstract = "Thioguanine-resistant primary clones were grown from single cell suspensions obtained from dog and human kidneys by enzymatic digestion. In medium containing a relatively high concentration (10μg/ ml) of thioguanine, thioguanine-resistant primary clones arose from each source at frequencies ranging from 10-4 to 10-5. A reduction in total hypoxanthine uptake was found in the thioguanine-resistant primary clones which had developed in thioguanine medium, consistent with a reduction in hypoxanthine phosphoribosyltransferase activity. When these thioguanine-resistant primary clones were subsequently grown in the absence of thioguanine and assayed for the thioguanine-resistant phenotype and hypoxanthine phosphoribosyltransferase activity, it was found that most were now thioguanine-sensitive and yielded cell free extracts with substantial amounts of hypoxanthine phosphoribosyltransferase activity. In contrast, thioguanine-resistant human clones grown continuously in the presence of thioguanine yielded cell free extracts with little or no detectable hypoxanthine phosphoribosyltransferase activity. Southern blot analysis demonstrated no structural alterations in the hypoxanthine phosphoribosyltransferase gene in thioguanine-resistant primary human kidney clones. These results suggest that a novel mechanism(s) for thioguanine resistance and the control of hypoxanth phosphoribosyltransferase expression may occur in dog and human kidney cells.",
keywords = "hypoxanthine phosphoribosyltransferase, kidney primary clones, purine analogue resistance",
author = "Mitchell Turker and Monnat, {Raymond J.} and Fukuchi, {Ken Ichiro} and Johnston, {Patricia A.} and Ogburn, {Charles E.} and Weller, {Richard E.} and Park, {James F.} and Martin, {George M.}",
year = "1988",
month = "6",
doi = "10.1007/BF00119247",
language = "English (US)",
volume = "4",
pages = "211--223",
journal = "Cell Biology and Toxicology",
issn = "0742-2091",
publisher = "Springer Netherlands",
number = "2",

}

TY - JOUR

T1 - A novel class of unstable 6-thioguanine-resistant cells from dog and human kidneys

AU - Turker, Mitchell

AU - Monnat, Raymond J.

AU - Fukuchi, Ken Ichiro

AU - Johnston, Patricia A.

AU - Ogburn, Charles E.

AU - Weller, Richard E.

AU - Park, James F.

AU - Martin, George M.

PY - 1988/6

Y1 - 1988/6

N2 - Thioguanine-resistant primary clones were grown from single cell suspensions obtained from dog and human kidneys by enzymatic digestion. In medium containing a relatively high concentration (10μg/ ml) of thioguanine, thioguanine-resistant primary clones arose from each source at frequencies ranging from 10-4 to 10-5. A reduction in total hypoxanthine uptake was found in the thioguanine-resistant primary clones which had developed in thioguanine medium, consistent with a reduction in hypoxanthine phosphoribosyltransferase activity. When these thioguanine-resistant primary clones were subsequently grown in the absence of thioguanine and assayed for the thioguanine-resistant phenotype and hypoxanthine phosphoribosyltransferase activity, it was found that most were now thioguanine-sensitive and yielded cell free extracts with substantial amounts of hypoxanthine phosphoribosyltransferase activity. In contrast, thioguanine-resistant human clones grown continuously in the presence of thioguanine yielded cell free extracts with little or no detectable hypoxanthine phosphoribosyltransferase activity. Southern blot analysis demonstrated no structural alterations in the hypoxanthine phosphoribosyltransferase gene in thioguanine-resistant primary human kidney clones. These results suggest that a novel mechanism(s) for thioguanine resistance and the control of hypoxanth phosphoribosyltransferase expression may occur in dog and human kidney cells.

AB - Thioguanine-resistant primary clones were grown from single cell suspensions obtained from dog and human kidneys by enzymatic digestion. In medium containing a relatively high concentration (10μg/ ml) of thioguanine, thioguanine-resistant primary clones arose from each source at frequencies ranging from 10-4 to 10-5. A reduction in total hypoxanthine uptake was found in the thioguanine-resistant primary clones which had developed in thioguanine medium, consistent with a reduction in hypoxanthine phosphoribosyltransferase activity. When these thioguanine-resistant primary clones were subsequently grown in the absence of thioguanine and assayed for the thioguanine-resistant phenotype and hypoxanthine phosphoribosyltransferase activity, it was found that most were now thioguanine-sensitive and yielded cell free extracts with substantial amounts of hypoxanthine phosphoribosyltransferase activity. In contrast, thioguanine-resistant human clones grown continuously in the presence of thioguanine yielded cell free extracts with little or no detectable hypoxanthine phosphoribosyltransferase activity. Southern blot analysis demonstrated no structural alterations in the hypoxanthine phosphoribosyltransferase gene in thioguanine-resistant primary human kidney clones. These results suggest that a novel mechanism(s) for thioguanine resistance and the control of hypoxanth phosphoribosyltransferase expression may occur in dog and human kidney cells.

KW - hypoxanthine phosphoribosyltransferase

KW - kidney primary clones

KW - purine analogue resistance

UR - http://www.scopus.com/inward/record.url?scp=0023727819&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0023727819&partnerID=8YFLogxK

U2 - 10.1007/BF00119247

DO - 10.1007/BF00119247

M3 - Article

VL - 4

SP - 211

EP - 223

JO - Cell Biology and Toxicology

JF - Cell Biology and Toxicology

SN - 0742-2091

IS - 2

ER -