Expressed antibody repertoires in human cord blood cells: 454 sequencing and IMGT/HighV-QUEST analysis of germline gene usage, junctional diversity, and somatic mutations.

Human cord blood cell-derived IgM antibodies are important for the neonate immune responses and construction of germline-based immunoglobulin libraries. Several previous studies of a relatively small number of sequences found that they exhibit restrictions in the usage of germline genes and in the diversity of the variable heavy chain complementarity determining region 3 compared to adults. To further characterize such restrictions on a larger scale and to compare the early B-cell diversity to adult IgM repertoires, we performed 454 sequencing and IMGT/HighV-QUEST analysis of cord blood IG libraries from two babies and determined germline gene usage, V-D-J rearrangement, VHCDR3 diversity, and somatic mutations to characterize human neonate repertoire. Most of the germline subgroups were identified with frequencies comparable to those present in the adult IgM repertoire except for the IGHV1-2 gene that was preferentially expressed in the cord blood cells. The gene usage diversity contributed to 1,430 unique IGH V-D-J rearrangement patterns while the exonuclease trimming and N region addition at the V-D-J junctions along with gene diversity created a wide range of VHCDR3 with different lengths and sequence variability. We observed a lower degree of somatic mutations in the CDR and framework regions of antibodies from cord blood cells compared to adults. These results provide insights into the characteristics of human cord blood antibody repertoires, which have gene usage diversity and VHCDR3 lengths similar to that of the adult IgM repertoire but differ significantly in some of the gene usages, V-D-J rearrangements, junctional diversity, and somatic mutations.

Classification of protein-DNA complexes based on structural descriptors.

We attempt to classify protein-DNA complexes by using a set of 11 descriptors, mainly characterizing protein-DNA interactions, including the number of atomic contacts at major and minor grooves, conformational deviations from standard B- and A-DNA forms, widths of DNA grooves, GC content, specificity measures of direct and indirect readouts, and buried surface area at the complex interface. The cluster analyses were carried out for a unique set of 62 complexes including a variety of protein motifs, and 7 distinct clusters were revealed from the analyses. We found that some proteins with the same motif are classified into different clusters, whereas different proteins with distinct motifs are classified into the same cluster. These results suggest that the conventional motif-based classification of DNA binding proteins may not necessarily correspond to structural and functional properties of protein-DNA complexes, and that the present classification will help to identify common properties and rules that govern protein-DNA recognition.