Lectin-resistant CHO cells: selection of new mutant phenotypes

Area of Science:

• Cell Biology
• Glycobiology
• Biochemistry

Background:

• Cytotoxic plant lectins are used to select for cell mutants with altered surface carbohydrates, indicating defects in glycosylation.
• Single lectin selections yield only the most common mutants due to limited specificity.
• Previous work demonstrated that combining lectins enhances selection specificity.

Purpose of the Study:

• To exploit combined lectin selection strategies for identifying novel lectin-resistant (LecR) mutants.
• To characterize the genetic basis of newly identified LecR phenotypes in Chinese hamster ovary (CHO) cells.

Main Methods:

• Utilized a combination of cytotoxic plant lectins for enhanced selection of LecR mutants.
• Performed genetic analysis using somatic cell hybridization to determine complementation groups of new mutants.
• Characterized five novel LecR phenotypes.

Main Results:

• Uncovered five new LecR phenotypes in CHO cells.
• One new phenotype represents a novel recessive complementation group.
• Two phenotypes exhibit dominant behavior in somatic cell hybrids, and two fall into previously identified complementation groups.

Conclusions:

• Combined lectin selection is an effective strategy for discovering novel glycosylation mutants.
• The identification of new complementation groups expands our understanding of glycosylation pathways.
• These novel LecR mutants provide valuable tools for studying specific defects in cellular glycosylation reactions.