Increased ribonucleotide reductase activity in hydroxyurea-resistant mosquito cells

• Published in: Archives of insect biochemistry and physiology Vol. 46; no. 1-2; pp. 19 - 25
• Main Authors: Gerenday, Anna, Shih, Karen M., Herman, Carter C., Fallon, Ann M.
• Format: Journal Article
• Language: English
• Published: New York John Wiley & Sons, Inc 01-01-2001
• Subjects: Aedes - enzymology; Aedes - physiology; Aedes albopictus; Animals; Cell Culture Techniques - methods; Culicidae; Drug Resistance - genetics; Enzyme Inhibitors - pharmacology; Hydroxyurea - pharmacology; hydroxyurea-resistant cell line; Karyotyping; Protein Biosynthesis; Proteins - analysis; ribonucleotide; Ribonucleotide Reductases - metabolism; Selection, Genetic
• ISSN: 0739-4462; 1520-6327
• DOI: 10.1002/arch.4

Hydroxyurea‐resistant Aedes albopictus mosquito cells were selected by incremental exposure of unmutagenized cells to hydroxyurea concentrations ranging from 0.1 to 8 mM. Clonal populations that had become 40‐fold more resistant to hydroxyurea than wild‐type cells varied in morphology, and their growth rate decreased to a ˜45 h doubling time, relative to an 18 h doubling time in unselected cells. At this level of resistance, the cells remained diploid, with a modal chromosome number of 6. When labelled with 35S[methionine/cysteine], clone HU1062, which grew in the presence of 8 mM hydroxyurea, overproduced a labeled protein with the approximate size of the 45,000 dalton M2 subunit of ribonucleotide reductase. Consistent with this observation, ribonucleotide reductase activity in HU‐1062 cells was approximately 10‐fold higher than in wild‐type control cells. This is the first example of an hydroxyurea‐resistant insect cell line. Arch. Insect Biochem. Physiol. 34:31–41, 1997. © 1997 Wiley‐Liss, Inc.