The C. elegans rab family: identification, classification and toolkit construction.

Area of Science:

• Molecular Biology
• Genomics
• Cell Biology

Background:

• Rab GTPases are crucial regulators of eukaryotic vesicle transport.
• Rab protein localization and function are often conserved across species.
• Model organisms like C. elegans offer insights into human diseases linked to Rab and vesicle trafficking defects.

Purpose of the Study:

• To identify and classify the complete Rab protein family in C. elegans.
• To develop a molecular toolset for studying Rab protein function in vesicle transport.

Main Methods:

• Bioinformatic analysis and molecular phylogenetics to classify Rab proteins.
• Comparative analysis of intron positions across species for difficult-to-classify genes.
• Utilized Gateway-compatible C. elegans ORFeome clones to create dominant-negative and constitutive active Rab ORFs.

Main Results:

• Successfully classified 30 out of 31 identified C. elegans Rab proteins.
• Identified Rab31/Rab50 as a potential member of the last eukaryotic common ancestor (LECA).
• Created a toolset of 44 full-length, sequence-verified Rab open reading frames (ORFs), including dominant-negative and constitutive active forms.

Conclusions:

• The study provides a comprehensive classification of the C. elegans Rab family.
• The developed molecular toolset facilitates research on Rab-mediated vesicle trafficking.
• This work supports the use of C. elegans as a model for understanding human diseases related to vesicle transport.