Increased ribonucleotide reductase activity in hydroxyurea‐resistant mosquito cells

Gerenday, Anna; Shih, Karen M.; Herman, Carter C.; Fallon, Ann M.

doi:10.1002/(sici)1520-6327(1997)34:1<31::aid-arch3>3.3.co;2-i

Cited by 2 publications

(4 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Only at the highest levels of hydroxyurea resistance have changes in the R1 protein and/or its corresponding gene been observed (reviewed by Wright, 1989). Consistent with these observations for mammalian cells, clonal populations of mosquito ( Aedes albopictus ) cells that were fortyfold more resistant to hydroxyurea than wild‐type cells showed increased levels of R2 protein and RNR enzyme activity ( Gerenday et al ., 1997b ). In the present paper, we describe the use of a PCR‐based approach to obtain the cDNA encoding the Ae.…”

Section: Introductionmentioning

confidence: 58%

“…To ensure accuracy, three individually derived pMosR2–5′ and pMosR2–3′ clones were sequenced. In addition, one RT PCR‐derived MosR2 cDNA from C7–10 cells, and five RT PCR‐derived MosR2 cDNAs from each of two hydroxyurea‐resistant cell lines, HU‐1062–10 and HU‐1062–15 ( Gerenday et al ., 1997b ) were cloned, sequenced, and found to be identical. This result indic ‐ ated that the increased R2 activity in hydroxyurea‐resistant mosquito cells was not due to a mutation in the coding region that enhanced enzyme activity.…”

Section: Resultsmentioning

confidence: 99%

“…The Ae. albopictus mosquito cells were from the phenotypically wild‐type clone C7–10 ( Fallon & Stollar, 1987) and from the hydroxyurea‐resistant clone HU‐1062 ( Gerenday et al ., 1997b ). Cells were maintained in E‐5 medium containing 5% foetal bovine serum, essentially as described previously.…”

Section: Resultsmentioning

confidence: 99%

See 2 more Smart Citations

Sequence of a mosquito ribonucleotide reductase cDNA and evidence for gene amplification in hydroxyurea‐resistant cells

Shotkoski

Jayachandran

Stamatoyannopoulos

et al. 1999

Insect Molecular Biology

View full text Add to dashboard Cite

Ribonucleotide reductase is essential for production of the deoxyribonucleotide substrates required for DNA synthesis in all eukaryotic cells. The full-length cDNA encoding a mosquito ribonucleotide reductase R2 subunit was obtained from Aedes albopictus cells using a polymerase chain reaction (PCR)-based approach. The cDNA contained 1197 nucleotides, and encoded a 398 amino acid R2 protein. Overall, mosquito R2 shared approximately 70% amino acid sequence identity with R2 proteins from vertebrates, but conservation at the N-terminus of the protein was relatively low. The sequence of R2 cDNA was identical in hydroxyurea-resistant mosquito cells and in wild-type cells, while the R2 gene copy number was increased ten to twentyfold in resistant cells.

show abstract

Section: Introductionmentioning

confidence: 58%

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Sequence of a mosquito ribonucleotide reductase cDNA and evidence for gene amplification in hydroxyurea‐resistant cells

Shotkoski

Jayachandran

Stamatoyannopoulos

et al. 1999

Insect Molecular Biology

View full text Add to dashboard Cite

show abstract

“…We have recently demonstrated that relatively low concentrations of hydroxyurea inhibit growth of wild type Aedes albopictus mosquito cells, and that the likely target for hydroyurea toxicity is ribonucleotide reductase (Gerenday et a/., 1997), a rate-limiting enzyme required for DNA synthesis. In this paper we describe synchronization of mosquito cells using single or double treatments with hydroxyurea.…”

Section: Introductionmentioning

confidence: 99%

Synchronization of Aedes albopictus mosquito cells using hydroxyurea

Gerenday¹,

Blauwkamp²,

Fallen³

1997

Insect Molecular Biology

View full text Add to dashboard Cite

We have established conditions for use of hydroxyurea, a reversible inhibitor of DNA synthesis, to synchronize the division cycle of a continuous cell line from the mosquito, Aedes albopictus. In the range of 0.15-0.25 mM hydroxyurea, an 18 h treatment, followed by removal of the drug, results in effective synchronization. When combined with the partial synchronization that occurs within 10 h of dilution and plating, more than 80% of cells treated with hydroxyurea could be recovered in the synthesis (S) phase of the cell cycle during the 4 h period after removal of the drug. The degree of synchrony was enhanced when cells were exposed to two consecutive hydroxyurea treatments spaced 10 h apart. Synchronized cells expressed maximal levels of a reporter gene when transfected immediately after removal of hydroxyurea. This is the first description of effective chemical synchronization of an insect cell line using hydroxyurea.

show abstract

Increased ribonucleotide reductase activity in hydroxyurea‐resistant mosquito cells

Cited by 2 publications

References 0 publications

Sequence of a mosquito ribonucleotide reductase cDNA and evidence for gene amplification in hydroxyurea‐resistant cells

Sequence of a mosquito ribonucleotide reductase cDNA and evidence for gene amplification in hydroxyurea‐resistant cells

Synchronization of Aedes albopictus mosquito cells using hydroxyurea

Contact Info

Product

Resources

About