Selection and characterization of chinese hamster ovary cells resistant to the cytotoxicity of lectins

Pamela Stanley, Louis Siminovitch

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Chinese hamster ovary (CHO) cells selected in a single step for resistance to the cytotoxicity of the lectin from red kidney beans (PHA) behave as authentic somatic cell mutants. The PHA-resistant (PhaR) phenotype is stable in the absence of selection; its frequency in a sensitive population is increased several-fold by mutagenesis; and it behaves recessively in somatic cell hybrids. The activity of a specific glycosyl transferase which transfers N-acetylglucosamine (GlcNAc) to terminal α-mannose residues is dramatically reduced (≤5% of the activity detected in wild-type CHO cells) in several independent PhaR clones. These clones also exhibit (a) a decreased ability to bind [125I]-PHA; (b) a marked resistance to the cytotoxicity of wheat germ agglutinin (WGA), Ricin (RIC) and Lens culinaris agglutinin (LCA); (c) a 4- to 5-fold increased sensitivity to the cytotoxicity of concanavalin A (Con A); (d) an increased ability to bind125I-Con A; and (e) decreased surface galactose residues-all properties consistent with the specific loss of the GlcNAc transferase activity. The lectins WGA, RIC, LCA and Con A have also been used to select, in a single step, resistant clones from each of two complementary CHO auxotrophic lines. These lectin-resistant clones have been characterized by their ability to survive cytotoxic doses of PHA, Con A, WGA, RIC or LCA, and 4-5 "lectin-resistance" phenotypes have been demonstrated. Complementation data is being sought by somatic cell hybridization. Preliminary results show that two phenotypically-distinct Con AR mutants are complementary in that hybrid cells formed between them exhibit wild-type sensitivity to Con A.

Original languageEnglish (US)
Pages (from-to)208-215
Number of pages8
JournalIn Vitro
Volume12
Issue number3
DOIs
StatePublished - Mar 1976
Externally publishedYes

Fingerprint

Cytotoxicity
Concanavalin A
Cricetulus
Ricin
Lectins
Wheat Germ Agglutinins
Ovary
Clone Cells
Cells
Hybrid Cells
Phenotype
Mutagenesis
Acetylglucosamine
Phytohemagglutinins
Vulnerable Populations
Mannose
Transferases
Galactose
lentil lectin

Keywords

  • complementation
  • glycosyl transferases
  • lectins
  • membrane mutants
  • somatic cell genetics

ASJC Scopus subject areas

  • Developmental Biology
  • Clinical Biochemistry
  • Cell Biology

Cite this

Selection and characterization of chinese hamster ovary cells resistant to the cytotoxicity of lectins. / Stanley, Pamela; Siminovitch, Louis.

In: In Vitro, Vol. 12, No. 3, 03.1976, p. 208-215.

Research output: Contribution to journalArticle

@article{56518aac29cb45489dc0a96844f33153,
title = "Selection and characterization of chinese hamster ovary cells resistant to the cytotoxicity of lectins",
abstract = "Chinese hamster ovary (CHO) cells selected in a single step for resistance to the cytotoxicity of the lectin from red kidney beans (PHA) behave as authentic somatic cell mutants. The PHA-resistant (PhaR) phenotype is stable in the absence of selection; its frequency in a sensitive population is increased several-fold by mutagenesis; and it behaves recessively in somatic cell hybrids. The activity of a specific glycosyl transferase which transfers N-acetylglucosamine (GlcNAc) to terminal α-mannose residues is dramatically reduced (≤5{\%} of the activity detected in wild-type CHO cells) in several independent PhaR clones. These clones also exhibit (a) a decreased ability to bind [125I]-PHA; (b) a marked resistance to the cytotoxicity of wheat germ agglutinin (WGA), Ricin (RIC) and Lens culinaris agglutinin (LCA); (c) a 4- to 5-fold increased sensitivity to the cytotoxicity of concanavalin A (Con A); (d) an increased ability to bind125I-Con A; and (e) decreased surface galactose residues-all properties consistent with the specific loss of the GlcNAc transferase activity. The lectins WGA, RIC, LCA and Con A have also been used to select, in a single step, resistant clones from each of two complementary CHO auxotrophic lines. These lectin-resistant clones have been characterized by their ability to survive cytotoxic doses of PHA, Con A, WGA, RIC or LCA, and 4-5 {"}lectin-resistance{"} phenotypes have been demonstrated. Complementation data is being sought by somatic cell hybridization. Preliminary results show that two phenotypically-distinct Con AR mutants are complementary in that hybrid cells formed between them exhibit wild-type sensitivity to Con A.",
keywords = "complementation, glycosyl transferases, lectins, membrane mutants, somatic cell genetics",
author = "Pamela Stanley and Louis Siminovitch",
year = "1976",
month = "3",
doi = "10.1007/BF02796443",
language = "English (US)",
volume = "12",
pages = "208--215",
journal = "In Vitro Cellular and Developmental Biology - Plant",
issn = "1054-5476",
publisher = "Springer New York",
number = "3",

}

TY - JOUR

T1 - Selection and characterization of chinese hamster ovary cells resistant to the cytotoxicity of lectins

AU - Stanley, Pamela

AU - Siminovitch, Louis

PY - 1976/3

Y1 - 1976/3

N2 - Chinese hamster ovary (CHO) cells selected in a single step for resistance to the cytotoxicity of the lectin from red kidney beans (PHA) behave as authentic somatic cell mutants. The PHA-resistant (PhaR) phenotype is stable in the absence of selection; its frequency in a sensitive population is increased several-fold by mutagenesis; and it behaves recessively in somatic cell hybrids. The activity of a specific glycosyl transferase which transfers N-acetylglucosamine (GlcNAc) to terminal α-mannose residues is dramatically reduced (≤5% of the activity detected in wild-type CHO cells) in several independent PhaR clones. These clones also exhibit (a) a decreased ability to bind [125I]-PHA; (b) a marked resistance to the cytotoxicity of wheat germ agglutinin (WGA), Ricin (RIC) and Lens culinaris agglutinin (LCA); (c) a 4- to 5-fold increased sensitivity to the cytotoxicity of concanavalin A (Con A); (d) an increased ability to bind125I-Con A; and (e) decreased surface galactose residues-all properties consistent with the specific loss of the GlcNAc transferase activity. The lectins WGA, RIC, LCA and Con A have also been used to select, in a single step, resistant clones from each of two complementary CHO auxotrophic lines. These lectin-resistant clones have been characterized by their ability to survive cytotoxic doses of PHA, Con A, WGA, RIC or LCA, and 4-5 "lectin-resistance" phenotypes have been demonstrated. Complementation data is being sought by somatic cell hybridization. Preliminary results show that two phenotypically-distinct Con AR mutants are complementary in that hybrid cells formed between them exhibit wild-type sensitivity to Con A.

AB - Chinese hamster ovary (CHO) cells selected in a single step for resistance to the cytotoxicity of the lectin from red kidney beans (PHA) behave as authentic somatic cell mutants. The PHA-resistant (PhaR) phenotype is stable in the absence of selection; its frequency in a sensitive population is increased several-fold by mutagenesis; and it behaves recessively in somatic cell hybrids. The activity of a specific glycosyl transferase which transfers N-acetylglucosamine (GlcNAc) to terminal α-mannose residues is dramatically reduced (≤5% of the activity detected in wild-type CHO cells) in several independent PhaR clones. These clones also exhibit (a) a decreased ability to bind [125I]-PHA; (b) a marked resistance to the cytotoxicity of wheat germ agglutinin (WGA), Ricin (RIC) and Lens culinaris agglutinin (LCA); (c) a 4- to 5-fold increased sensitivity to the cytotoxicity of concanavalin A (Con A); (d) an increased ability to bind125I-Con A; and (e) decreased surface galactose residues-all properties consistent with the specific loss of the GlcNAc transferase activity. The lectins WGA, RIC, LCA and Con A have also been used to select, in a single step, resistant clones from each of two complementary CHO auxotrophic lines. These lectin-resistant clones have been characterized by their ability to survive cytotoxic doses of PHA, Con A, WGA, RIC or LCA, and 4-5 "lectin-resistance" phenotypes have been demonstrated. Complementation data is being sought by somatic cell hybridization. Preliminary results show that two phenotypically-distinct Con AR mutants are complementary in that hybrid cells formed between them exhibit wild-type sensitivity to Con A.

KW - complementation

KW - glycosyl transferases

KW - lectins

KW - membrane mutants

KW - somatic cell genetics

UR - http://www.scopus.com/inward/record.url?scp=0017064630&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0017064630&partnerID=8YFLogxK

U2 - 10.1007/BF02796443

DO - 10.1007/BF02796443

M3 - Article

C2 - 1262042

AN - SCOPUS:0017064630

VL - 12

SP - 208

EP - 215

JO - In Vitro Cellular and Developmental Biology - Plant

JF - In Vitro Cellular and Developmental Biology - Plant

SN - 1054-5476

IS - 3

ER -