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Molecular cloning of feline CC-chemokine cDNAs
Y Endo1, T Mizuno, Y Nishimura
1Department of Veterinary Internal Medicine, Graduate School of Agricultural and Life Sciences, University of Tokyo, Japan. ayendo@ecc-mail.hongo.ecc.u-tokyo.ac.jp
Abstract:
cDNA clones of feline chemokines, MIP-1alpha, MIP-1beta and RANTES, were molecularly isolated with the purpose of using these sequences for future investigation of the inhibitory effects on lentivirus entry and their role in immunological functions. The feline MIP-1alpha and MIP-1beta cDNA clones spanned their entire coding regions encoding 93 and 92 amino acids, respectively. The amino acid sequences of feline MIP-1alpha and MIP-1beta compared to those of their human, mouse and rat counterparts showed similarities of 75.3-79.6% and 73.9-88.0%, respectively. Feline MIP-1alpha and MIP-1beta had four conserved cysteines with a structure made up of the first two cysteines that are characteristic of the CC-chemokine subfamily. The amino terminal of these MIP-1alpha and MIP-1beta sequences was distinctly hydrophobic, suggesting that they may function as signal peptides. A partial cDNA clone consisting of 193 bp was obtained for feline RANTES, and it also showed a high degree of sequence similarity to those of other species and contained the characteristic structure made up of adjacent cysteines. These molecular clones of feline chemokines will be useful in the examination of their inhibitory effect on the cellular entry of feline immunodeficiency virus.
Insights
Researchers isolated feline chemokine cDNA clones, including MIP-1alpha, MIP-1beta, and RANTES. These sequences will aid in studying their role in immunity and inhibiting lentivirus entry, particularly feline immunodeficiency virus.
Area of Science:
- Immunology
- Molecular Biology
- Virology
Background:
- Chemokines are crucial signaling proteins in the immune system.
- Feline immunodeficiency virus (FIV) poses a significant threat to feline health.
- Understanding feline chemokine function is vital for developing antiviral strategies.
Purpose of the Study:
- To molecularly isolate cDNA clones of feline chemokines: MIP-1alpha, MIP-1beta, and RANTES.
- To investigate the potential of these chemokines in inhibiting lentivirus entry.
- To explore their role in feline immunological functions.
Main Methods:
- Molecular isolation of cDNA clones for feline MIP-1alpha, MIP-1beta, and RANTES.
- Sequencing and analysis of the isolated cDNA clones.
- Comparative sequence analysis with homologous chemokines from other species.
Main Results:
- Full-length coding regions for feline MIP-1alpha (93 amino acids) and MIP-1beta (92 amino acids) were obtained.
- High sequence similarity (75.3-88.0%) was observed between feline and other mammalian MIP-1alpha and MIP-1beta.
- Feline MIP-1alpha and MIP-1beta possess conserved cysteines characteristic of CC-chemokines and hydrophobic N-termini, suggesting signal peptide function.
- A partial cDNA clone for feline RANTES was isolated, also showing high sequence similarity and characteristic CC-chemokine structure.
- The isolated clones provide sequences for future studies on FIV inhibition.
Conclusions:
- The molecular clones of feline MIP-1alpha, MIP-1beta, and RANTES have been successfully generated.
- These clones are valuable tools for investigating the inhibitory effects of these chemokines on feline lentivirus entry.
- The findings contribute to understanding feline chemokine function and developing potential FIV therapies.