Different quaternary structures of human RECQ1 are associated with its dual enzymatic activity

RecQ helicases are essential for the maintenance of chromosome stability. In addition to DNA unwinding, some RecQ enzymes have an intrinsic DNA strand annealing activity. The function of this dual enzymatic activity and the mechanism that regulates it is, however, unknown. Here, we describe two quaternary forms of the human RECQ1 helicase, higher-order oligomers consistent with pentamers or hexamers, and smaller oligomers consistent with monomers or dimers. Size exclusion chromatography and transmission electron microscopy show that the equilibrium between the two assembly states is affected by single-stranded DNA (ssDNA) and ATP binding, where ATP or ATPγS favors the smaller oligomeric form. Our three-dimensional electron microscopy reconstructions of human RECQ1 reveal a complex cage-like structure of approximately 120 Å × 130 Å with a central pore. This oligomeric structure is stabilized under conditions in which RECQ1 is proficient in strand annealing. In contrast, competition experiments with the ATPase-deficient K119R and E220Q mutants indicate that RECQ1 monomers, or tight binding dimers, are required for DNA unwinding. Collectively, our findings suggest that higher-order oligomers are associated with DNA strand annealing, and lower-order oligomers with DNA unwinding.     

Comparing methods for estimating the abundance of Western Capercaillie Tetrao urogallus males in Pyrenean leks: singing counts versus genetic analysis of non-invasive samples

ABSTRACT

Population estimates of male Western Capercaillies Tetrao urogallus were carried out during the mating season using two methods: counts of singing males and non-invasive genetic analysis. Estimates of male numbers were 50% lower using the singing counts compared to the estimates obtained through genetic analysis, and underestimates were greatest when the number of Capercaillies was lowest.     

Size matters in quantitative radar monitoring of animal migration: estimating monitored volume from wingbeat frequency

Quantitative radar studies are an important component of studying the movements of birds. Whether a bird, at a certain distance from the radar, is detected or not depends on its size. The volume monitored by the radar is therefore different for birds of different sizes. Consequently, an accurate quantification of bird movements recorded by small‐scale radar requires an accurate determination of the monitored volume for the objects in question, although this has tended to be ignored. Here, we demonstrate the importance of sensitivity settings for echo detection on the estimated movement intensities of birds of different sizes. The amount of energy reflected from a bird and detected by the radar receiver (echo power) depends not only on the bird's size and on the distance from the radar antenna, but also on the beam shape and the bird's position within this beam. We propose a method to estimate the size of a bird based on the wingbeat frequency, retrieved from the echo‐signal, independent of the absolute echo power. The estimated bird‐size allows calculation of size‐specific monitored volumes, allowing accurate quantification of movement intensities. We further investigate the importance of applying size‐specific monitored volumes to quantify avian movements instead of using echo counts. We also highlight the importance of accounting for size‐specific monitored volume of small scale radar systems, and the necessity of reporting technical information on radar parameters. Applying this framework will increase the quality and validity of quantitative radar monitoring.     

Differential growth factor responses of epithelial cell cultures derived from normal human prostate,benign prostatic hyperplasia,and primary prostate carcinoma

Because of a lack of information of the optimum nutritional requirements, epithelial cells derived from normal human prostate and prostate tumors have been difficult to propagate in vitro, which hinders research in prostate carcinogenesis. In an effort to establish optimum nutritional conditions and differences in growth characteristics of normal human prostate (NP), benign prostatic hyperplasia (BPH), and prostatic carcinoma (PCA), we have compared the effects of several growth factors on cell proliferation and elucidated growth properties of low passage epithelial cells derived from NP, BPH, and PCA of an African-American patient. Primary and low passage cultures were propagated in serum-free keratinocyte basal medium (KBM) supplemented with insulin (5 μg/ml), hydrocortisone (0.5 μg/ml), epidermal growth factor (EGF, 10 ng/ml), bovine pituitary extract (BPE; 50 μg/ml), cholera toxin (10 ng/ml), and antibiotics. Almost all NP, BPH, and PCA cells were positive for cytokeratins and prostate-specific antigen (PSA). The NP, BPH, and PCA cells were essentially diploid and lacked mutations in c-K-ras and c-Ha-ras oncogenes, and p53 tumor suppressor gene. However, they exhibited progressively accelerating growth parameters. The population doubling times of NP, BPH and PCA were 51 hr, 37 hr, and 29 hr, respectively; their saturation densities were 2.9 × 10⁴/cm², 3.3 × 10⁴/cm², and 7.2 × 10⁴/cm², respectively. The NP and BPH cells required all of the growth factors in the medium, as deletion of any one of the above factors strongly inhibited their growth. The PCA cells, however, were independent of EGF and hydrocortisone. PC-3, an established human prostate cancer cell line, was independent of the growth factors tested. Fetal bovine serum (FBS) inhibited the growth of NP, BPH and PCA cells. In contrast, FBS stimulated the growth of the PC-3 cells in a concentration-dependent manner. These results indicate that in the absence of any apparent karyotype alterations and mutations in c-K-ras, c-Ha-ras and p53 genes, epithelial cells derived from NP, BPH, and PCA exhibit significant differences in their growth properties and responses to growth factors. These variations may represent early changes involved in prostate cancer, while gene mutations and cytogenetic alterations occur in advanced and/or metastatic tumors. © 1996 Wiley-Liss, Inc.