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Published on: September 11, 2022
Sequence, chromosomal location and expression analysis of the murine homologue of human RAD51L2/RAD51C
C S Leasure1, J Chandler, D J Gilbert
1Mouse Cancer Genetics Program, National Cancer Institute at Frederick, P.O. Box B, Frederick, MD 21702, USA.
Abstract:
The Rad51 protein has been shown to play a vital role in the DNA repair process. In humans, its interaction with proteins like BRCA1 and BRCA2 has provided an insight into the mechanism of how these molecules function as tumor suppressors. Several members of the Rad51-like family have been recently identified, including RAD51L2. This gene has been found to be amplified in breast tumors suggesting its role in tumor progression. Here, we describe the cloning of the murine homologue of the human RAD51L2/RAD51C gene. Sequence analysis has revealed that the murine Rad51l2 protein is 86% identical and 93% similar to its human homologue. In spite of such high sequence conservation, the murine protein lacks the first nine amino acids present in the human protein. We have cloned and confirmed the sequence of the 5' end of the murine Rad51l2 cDNA using 5' RACE technique as well as by sequencing the genomic region flanking the first exon of the murine Rad51l2 gene. Northern analysis shows that Rad51l2 is expressed in several adult tissues as well as in embryos at various developmental stages. The murine Rad51l2 gene maps to chromosome 11 and is located in the syntenic region of human chromosome 17q22-23, where the human RAD51L2 is present.
Insights
Researchers cloned the mouse Rad51l2 gene, crucial for DNA repair and tumor suppression. Despite high similarity to human RAD51L2, the mouse version lacks nine amino acids, impacting its function in breast cancer progression.
Area of Science:
- Molecular Biology
- Genetics
- Cancer Research
Background:
- The Rad51 protein family is essential for DNA repair mechanisms.
- Human RAD51L2 (also known as RAD51C) is implicated in tumor suppression, with gene amplification observed in breast tumors.
- Understanding homologous proteins across species aids in elucidating complex biological functions.
Purpose of the Study:
- To clone and characterize the murine homologue of the human RAD51L2/RAD51C gene.
- To investigate the sequence, expression, and chromosomal localization of the mouse Rad51l2 gene.
- To provide insights into the potential role of Rad51l2 in biological processes, including cancer.
Main Methods:
- Cloning of the murine Rad51l2 gene.
- Sequence analysis to compare murine and human homologues.
- 5' RACE (Rapid Amplification of cDNA Ends) technique to confirm the 5' end sequence.
- Genomic sequencing of the region flanking the first exon.
- Northern blot analysis for gene expression profiling.
- Chromosomal mapping of the murine gene.
Main Results:
- The murine Rad51l2 protein shares 86% identity and 93% similarity with its human counterpart.
- A notable difference is the absence of the first nine amino acids in the murine protein compared to the human protein.
- Rad51l2 expression was detected in various adult tissues and embryonic developmental stages.
- The murine Rad51l2 gene was mapped to chromosome 11, syntenic to the human RAD51L2 locus on chromosome 17q22-23.
Conclusions:
- The study successfully cloned and characterized the murine Rad51l2 gene, a homologue of human RAD51C.
- High sequence conservation suggests a conserved function, while the N-terminal difference may indicate functional divergence.
- The expression pattern and conserved syntenic location support the importance of Rad51l2 in mammalian biology and evolution.
