The formation of a complex between RecA protein and single-stranded (ss) DNA was studied systematically by atomic force microscopy (AFM) by varying incubation time and the molecular ratio of RecA protein to single-stranded DNA binding (SSB) protein. New intermediate structures, such as small circular, tangled, and protruded structures in the absence of SSB and sharply turned structures in the presence of SSB, were clearly identified at the early stage of complex formation. These structures have probably resulted from competitive binding of RecA and SSB to DNA. After long incubation, only fully covered RecA-ssDNA and totally RecA-free SSB-ssDNA complexes were present regardless of RecA concentrations. Together with intermediate structures which consisted of only two parts, that is, ssDNA covered by SSB and by RecA proteins, the observation suggested strong neighbor cooperative binding of RecA to ssDNA assisted by SSB.

Allosteric Regulation↗

PubMedDate not supplied

DNase I hypersensitive sites in the chromatin of human mu immunoglobulin heavy-chain genes.

An immunoglobulin polypeptide chain is encoded by multiple gene segments that lie far apart in germ-line DNA and must be brought together to allow expression of an immunoglobulin gene active in B lymphocytes. For the immunoglobulin heavy chain genes, one of many variable (V) region genes becomes joined to one of several diversity (D) segments which are fused to one of several joining (J) segments lying 5' of the constant region (C) genes. Here we show that the rearranged mu genes of an IgM-producing human B-lymphocyte cell line exhibit pancreatic deoxyribonuclease (DNase I) hypersensitive sites in the JH-C mu intron that are absent in naked DNA or the chromatin of other differentiated cell types. DNA sequence analysis reveals that the major hypersensitive site maps to a conserved region of the JH-C mu intron recently shown to function as a tissue-specific enhancer of heavy-chain gene expression. A similar association of an enhancer-like element with a DNase I hypersensitive site has been reported for the mouse immunoglobulin light-chain J kappa-C kappa intron. These results implicate disruption of local chromatin structure in the mechanism of immunoglobulin enhancer function.

B-Lymphocytes↗

PubMedDate not supplied

Chemical cross-linking of peptides derived from RecA with single-stranded oligonucleotides containing 5-formyl-2'-deoxyuridine.

We report the first example of chemical cross-linking of 5-formyl-2'-deoxyuridine containing oligonucleotides with oligopeptides through a Schiff base formation. Twenty amino acid residue peptides investigated here were derived from the DNA binding site of RecA protein. We have demonstrated that the lysine residue placed at the 6th or 8th position from the N-terminus of the peptide directly contacts with DNA.

Amino Acid Sequence↗

PubMedDate not supplied

Articular cartilage contact pressure after tibial tuberosity transfer. A cadaveric study.

Medial transfer of the tibial tuberosity has been commonly used for treatment of recurrent dislocation of the patella and patellofemoral malalignment. In this study, six fresh human cadaveric knees were used. Static intrajoint loads were recorded using Fuji Prescale pressure-sensitive film for contact pressure and contact area determination in a closed kinetic chain knee testing protocol. Peak pressures, average contact pressures, and contact areas of the patellofemoral and tibiofemoral joints were calculated on native intact knee specimens and after tibial tuberosity transfer. All native intact knee specimens had a normal Q angle. Medialization of the tibial tuberosity significantly increased the patellofemoral contact pressure. Medial displacement of the tibial tuberosity also significantly increased the average contact pressure of the medial tibiofemoral compartment and changed the balance of tibiofemoral joint loading. The results of our study suggest that caution should be used when transferring a patellar tendon in the face of a preexisting normal Q angle as this will result in abnormally high peak pressure within the tibiofemoral joint. Overmedialization of the tibial tuberosity should be avoided in the varus knee, the knee after medial meniscectomy, and the knee with preexisting degenerative arthritis of the medial compartment.

Adult↗