Protein Repair Enzyme Research Articles

Isolation and characterization of transcription signal sequences from Streptococcus thermophilus.

Streptococcus thermophilus (ST) chromosomal DNA fragments generated by partial Sau3A digestion were cloned into the unique BamHI site upstream from the promoterless chloramphenicol acetyltransferase (cat) gene of the Escherichia coli (EC)promoter-probe vector pKK520-3. Recombinant plasmids containing ST sequences with transcription-activation activity were isolated from chloramphenicol-resistant (CmR) EC transformants. A promoterless Streptomyces antibioticus melanin biosynthesis operon (melC) was inserted immediately downstream from the ST sequence to identify DNA with strong promoter activity. Several ST transcription-activation sequences, termed STPs, were isolated and subcloned, and their nucleic acid sequences determined. The -10 and -35 consensus sequences were identified in these putative ST promoters. Detailed analysis of STP3306 sequence data revealed two partial open reading frames (ORFs) with high degrees of homology to prokaryotic GTP-binding protein and DNA repair enzyme, thus providing valuable information for further study on DNA maintenance in this important lactic acid bacterium.

Exceptional Seed Longevity and Robust Growth: Ancient Sacred Lotus from China

A 1,288 ± 271-yr-old (1,350 ± 220 yr BP, radiocarbon age) seed of Sacred Lotus (Nelumbo nucifera Gaertn.) from an ancient lake bed at Pulantien, Liaoning Province, China, has been germinated and subsequently radiocarbon dated. This is the oldest demonstrably viable and directly dated seed ever reported, the preserved relict of one of the early crops of lotus cultivated by Buddhists at Pulantien after introduction of the religion into the region prior to 372 A.D. A small portion of the dry pericarp of a second lotus fruit from the same locale has been dated as being 332 ± 135-yr-old (270 ± 60 yr BP, radiocarbon age) by accelerator mass spectroscopy at the Lawrence Livermore National Laboratory. This polycentenarian seed not only germinated but is still growing (since March, 1994). Of six old lotus fruits tested, two-thirds germinated, almost all in fewer than 4 d, as rapidly as fruits harvested from the progeny of Pulantien Sacred Lotus plants (under cultivation by the National Park Service in Washington, DC), and more rapidly than fresh fruits of Yellow Lotus [N. lutea (Willd.) Pers.]. Growth of the old lotus is robust: rhizome formation and leaf emergence at rhizome nodes are more rapid than those of the Pulantien progeny, although the leaf width is smaller. Activity of the protein-repair enzyme L-isoaspartyl methyltransferase in the old lotus seed is persistent during germination and is as robust as that in the progeny, and the degree of aspartyl racemization in proteins of the two groups of plants is minimal and essentially identical. The six dated ancient Sacred Lotus fruits range in age from 95 to 1,288 yr (with a mean age of 595 ± 380 yr), evidently reflecting their production, deposition, and preservation at varying times during the intervening millennium.

Immunolocalization of heat shock proteins in ragged-red fibers of patients with mitochondrial encephalomyopathies

Monoclonal antibodies against the 60 kDa heat shock protein (HSP-60) and against ubiquitin (UB) were used to study the expression of these proteins in muscle samples from patients with qualitative and quantitative alterations of mitochondrial DNA (mtDNA). We found an enhanced expression of HSP-60 and UB that was preferentially localized in ragged-red fibers (RRFs). HSP-60 may act as a protein repair enzyme catalyzing the refolding of misfolded proteins in the matrix of mitochondria of RRFs. On the other hand, UB could promote the elimination of abnormal proteins by its covalent interaction to substrates.