The ribonuclease from an extinct bovid ruminant

Area of Science:

• Evolutionary biology
• Molecular biology
• Biochemistry

Background:

• Ribonucleases (RNases) are crucial enzymes involved in RNA metabolism.
• Understanding the evolutionary trajectory of proteins provides insights into molecular adaptation.
• The evolutionary history of ruminant RNases is not fully elucidated.

Purpose of the Study:

• To reconstruct the ancestral ribonuclease sequence of the ancestor of swamp buffalo, river buffalo, and ox.
• To synthesize and characterize laboratory versions of ancestral and intermediate ribonuclease sequences.
• To investigate the functional evolution of ribonuclease in ruminants.

Main Methods:

• Phylogenetic analysis using the 'maximum parsimony' method to infer the ancestral sequence.
• Site-directed mutagenesis to create synthetic gene sequences for ancestral and intermediate proteins.
• Biochemical assays to examine the properties and functions of the synthesized ribonuclease variants.

Main Results:

• The ribonuclease sequence from the ancestor of modern buffalo and ox (approximating Pachyportax latidens) was computationally reconstructed.
• Synthetic ancestral and intermediate ribonuclease proteins were successfully generated in the laboratory.
• The functional properties of these reconstructed proteins were experimentally evaluated.

Conclusions:

• The study provides a reconstructed ancestral ribonuclease sequence for key ruminant species.
• Laboratory synthesis and characterization of ancestral proteins enable direct study of evolutionary changes.
• This research offers insights into the molecular evolution of ribonuclease in the Bovidae family.