SUMOylation regulates nuclear localization of Krüppel-like factor 5

SUMOylation is a form of post-translational modification shown to control nuclear transport. Krüppel-like factor 5 (KLF5) is an important mediator of cell proliferation and is primarily localized to the nucleus. Here we show that mouse KLF5 is SUMOylated at lysine residues 151 and 202. Mutation of these two lysines or two conserved nearby glutamates results in the loss of SUMOylation and increased cytoplasmic distribution of KLF5, suggesting that SUMOylation enhances nuclear localization of KLF5. Lysine 151 is adjacent to a nuclear export signal (NES) that resembles a consensus NES. The NES in KLF5 directs a fused green fluorescence protein to the cytoplasm, binds the nuclear export receptor CRM1, and is inhibited by leptomycin and site-directed mutagenesis. SUMOylation facilitates nuclear localization of KLF5 by inhibiting this NES activity, and enhances the ability of KLF5 to stimulate anchorage-independent growth of HCT116 colon cancer cells. A survey of proteins whose nuclear localization is regulated by SUMOylation reveals that SUMOylation sites are frequently located in close proximity to NESs. A relatively common mechanism for SUMOylation to regulate nucleocytoplasmic transport may lie in the interplay between neighboring NES and SUMOylation motifs.     

RAD51C interacts with RAD51B and is central to a larger protein complex in vivo exclusive of RAD51.

RAD51B and RAD51C are two of five known paralogs of the human RAD51 protein that are thought to function in both homologous recombination and DNA double-strand break repair. This work describes the in vitro and in vivo identification of the RAD51B/RAD51C heterocomplex. The RAD51B/RAD51C heterocomplex was isolated and purified by immunoaffinity chromatography from insect cells co-expressing the recombinant proteins. Moreover, co-immunoprecipitation of the RAD51B and RAD51C proteins from HeLa, MCF10A, and MCF7 cells strongly suggests the existence of an endogenous RAD51B/RAD51C heterocomplex. We extended these observations to examine the interaction between the RAD51B/RAD51C complex and the other RAD51 paralogs. Immunoprecipitation using protein-specific antibodies showed that RAD51C is central to a single large protein complex and/or several smaller complexes with RAD51B, RAD51D, XRCC2, and XRCC3. However, our experiments showed no evidence for the inclusion of RAD51 within these complexes. Further analysis is required to elucidate the function of the RAD51B/RAD51C heterocomplex and its association with the other RAD51 paralogs in the processes of homologous recombination and DNA double-strand break repair.     

Polymorphic microsatellite loci from the Southeast Asian cyprinid,Barbodes gonionotus (Bleeker)

The cyprinid Barbodes gonionotus (Bleeker) is a commercially important fish in both capture fisheries and aquaculture in Southeast Asia. Five polymorphic microsatellite loci from B. gonionotus are described. Four are highly variable, with 9–30 alleles observed per locus in four populations sampled from Thailand (H_O = 0.694–0.808). These will be of use in studies of population genetic structure and in pedigree analyses.     

Geology of the Early Arikareean Sharps Formation on the Pine Ridge Indian Reservation and Surrounding Areas of South Dakota and Nebraska

Based on geologic mapping, measured sections, and lithologic correlations, the local features of the upper and lower type areas of the Early Arikareean (30.8–20.6 million years ago) Sharps Formation are revised and correlated. The Sharps Formation above the basal Rockyford Member is divided into two members of distinct lithotypes. The upper 233 feet of massive siltstones and sandy siltstones is named the Gooseneck Road Member. The middle member, 161 feet of eolian volcaniclastic siltstones with fluvially reworked volcaniclastic lenses and sandy siltstone sheets, is named the Wolff Camp Member. An ashey zone at the base of the Sharps Formation is described and defined as the Rockyford Ash Zone (RAZ) in the same stratigraphic position as the Nonpareil Ash Zone (NPAZ) in Nebraska. Widespread marker beds of fresh water limestones at 130 feet above the base of the Sharps Formation and a widespread reddish-brown clayey siltstone at 165 feet above the base of the Sharps Formation are described. The Brown Siltstone Beds of Nebraska are shown to be a southern correlative of the Wolff Camp Member and the Rockyford Member of the Sharps Formation. Early attempts to correlate strata in the Great Plains were slow in developing. Recognition of the implications of the paleomagnetic and lithologic correlations of this paper will provide an added datum assisting researchers in future biostratigraphic studies. Based on similar lithologies, the Sharps Formation, currently assigned to the Arikaree Group, should be reassigned to the White River Group.