Detection of multiple, novel reverse transcriptase coding sequences in human nucleic acids: relation to primate...

Area of Science:

• Molecular Biology
• Genomics
• Retroviral Research

Background:

• Reverse transcriptase (RT) is a key enzyme in retroviral replication.
• Understanding RT coding sequences in host genomes is crucial for retroviral research.
• Human and mouse genomes may harbor diverse endogenous retroviral sequences.

Purpose of the Study:

• To detect and characterize novel reverse transcriptase coding sequences in human and mouse DNA.
• To compare the efficiency of different molecular techniques for sequence detection.
• To investigate the relationship of detected sequences to known retroviruses and endogenous elements.

Main Methods:

• Chemically synthesized oligonucleotides based on amino acid and nucleotide sequence data.
• Southern blotting, library screening, and polymerase chain reaction (PCR).
• PCR using consensus and unique oligonucleotide primers targeting RT sequences.

Main Results:

• A large number of novel reverse transcriptase coding sequences were detected in human and mouse DNA.
• PCR was the most rapid and productive method for sequence detection.
• Human DNA contains diverse retrovirus-related sequences, including those related to human T-cell leukemia virus (HTLV) types I and II, L-1 elements, and endogenous proviruses.
• HTLV type I-related sequences are transcribed in normal human T cells and a teratocarcinoma cell line.

Conclusions:

• Human and mouse genomes harbor a wide spectrum of retrovirus-related reverse transcriptase coding sequences.
• The polymerase chain reaction is a highly effective tool for identifying these sequences.
• Endogenous retroviral sequences, including those related to HTLV, are present and potentially active in the human genome.