Archaeal chromatin proteins histone HMtB and Alba have lost DNA-binding ability in laboratory strains of Methanothermobacter thermautotrophicus.
Alignments of the sequences of the all members of the archaeal histone and Alba1 families of chromatin proteins identified isoleucine residues, I19 in HMtB and I39 in MtAlba, in Methanothermobacter thermautotrophicus, at locations predicted to be directly involved in DNA binding. In all other HMfB family members, residue 19 is an arginine (R19), and either arginine or lysine is present in almost all other Alba1 family members at the structural site equivalent to I39 in MtAlba. Electrophoretic mobility shift assays revealed that recombinant HMtB and MtAlba do not bind DNA, but variants constructed with R19 and R39, respectively, bound DNA; and whereas MtAlba(I19) did not bind RNA, MtAlba(R19) bound both single stranded RNA and tRNA. Amplification and sequencing of MT0254 (encodes HMtB) and MT1483 (encodes MtAlba) from several Methanothermobacter thermautotrophicus lineages has revealed that HMtB and MtAlba had arginine residues at positions 19 and 39, respectively, in the original isolate and that spontaneous mutations must have occurred, and been fixed, in some laboratory lineages that now have HMtB(I19) and MtAlba(I39). The retention of these variants suggests some continuing functions and fusion of the HMtB(I19) sequence to HMtA2 resulted in a protein that folds to form a histone fold heterodimer that binds and compacts DNA. The loss of DNA binding by HMtB(I19) does not therefore prevent HMtB from participating in DNA interactions as one partner of an archaeal histone heterodimer.
Sandman
Kathleen
K
Department of Microbiology, Ohio State University, Columbus, OH 43210, USA.
Louvel
Hélène
H
Samson
Rachel Y
RY
Pereira
Suzette L
SL
Reeve
John N
JN
D019843:Archaeal Proteins / Q000378:metabolism*; D001483:Base Sequence; D017931:DNA Primers; D019641:DNA, Archaeal / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D019281:Dimerization; D004587:Electrophoresis, Agar Gel; D024202:Electrophoretic Mobility Shift Assay; D017013:Methanobacteriaceae / Q000254:growth & development / Q000378:metabolism*; D010957:Plasmids; D011485:Protein Binding; D011994:Recombinant Proteins / Q000378:metabolism
D019843:Archaeal Proteins; D017931:DNA Primers; D019641:DNA, Archaeal; D004268:DNA-Binding Proteins; D011994:Recombinant Proteins; C078607:HmtB protein, Methanothermobacter thermautotrophicus