A chromatin binding site in the tail domain of nuclear lamins that interacts with core histones.

Abstract

Abstract. Interaction of chromatin with the nuclear envelope and lamina is thought to help determine higher order chromosome organization in the interphase nucleus. Previous studies have shown that nuclear lamins bind chromatin directly. Here we have localized a chromatin binding site to the carboxyl-terminal tail domains of both A-and B-type mammalian lamins, and have characterized the biochemical properties of this binding in detail. Recombinant glutathione-S-transferase fusion proteins containing the tail domains of mammalian lamins C, BI, and B 2 were analyzed for their ability to associate with rat liver chromatin fragments immobilized on microtiter plate wells. We found that all three lamin tails specifically bind to chromatin with apparent Kds of 120-300 nM. By examining a series of deletion mutants, we have mapped the chromatin binding region of the lamin C tail to amino acids 396--430, a segment immediately adjacent to the rod domain. Furthermore, by analysis of chromatin subfractions, we found that core histones constitute the principal chromatin binding component for the lamin C tail. Through cooperativity, this lamin-histone interaction could be involved in specifying the high avidity attachment of chromatin to the nuclear envelope in vivo.T HE nuclear lamina is a filamentous protein meshwork that lines the nucleoplasmic surface of the nuclear envelope (NE) 1 (reviewed by Gerace and Burke, 1988;McKeon, 1991;Nigg, 1992;Georgatos et al., 1994). The lamina is thought to provide a structural framework for the NE and an anchoring site at the nuclear periphery for interphase chromosomes, and therefore could play a major role in interphase nuclear organization. The lamina consists of a polymeric assembly of nuclear lamins, members of the intermediate filament (IF) protein superfamily (see McKeon, 1991;Nigg, 1992), as well as a number of less abundant lamina-associated polypeptides (discussed by Gerace and Foisner, 1994). Vertebrate lamins are classified as A-or B-subtypes based on their sequence and biochemical properties. B-type lamins (lamins B1 and B2) are present in somatic cells throughout development, while A-type lamins (lamins A and C) are expressed only during or after terminal differentiation in most cells. Mam-

Conformational Changes in the Nuclear Lamina Induced by Herpes Simplex Virus Type 1 Require Genes U L 31 and U L 34

“…Less chromatin in regions of the nucleus could conceivably enhance exposure of the nucleoplasmic face of the inner nuclear membrane and thereby facilitate the egress of nucleocapsids. Since amino acids 411 to 553 of lamin A/C comprise a globular DNA binding domain that helps mediate association with chromatin (5,13,17,28,30), it was of interest that, through epitope mapping and GST pulldown reactions, we have determined that amino acids in or near the lamin A tail domain (amino acids 369 to 519 and 490 to 660) are sufficient to interact with U L 31 protein.…”

Aging of Hutchinson–Gilford progeria syndrome fibroblasts is characterised by hyperproliferation and increased apoptosis

“…Furthermore, disruption of normal lamin organization by microinjection of a dominant-negative lamin A mutant results in inhibition of both DNA replication (Spann et al, 1997) and RNA polymerase II-dependent transcription (Spann et al, 2002) suggesting a role for lamin A in both these processes. Finally, lamins have the ability to bind directly to DNA (Stierle et al, 2003) and to chromatin (Glass et al, 1993;Taniura et al, 1995) and indirectly via associations with proteins containing a LEM box (Lee et al, 2001;Martins et al, 2003). Thus, the list of possible functions of the A-type lamins includes maintenance of nuclear structural integrity, organisation of higher-order chromatin structure and control of gene expression (Hutchison, 2002).…”

Essential Roles forCaenorhabditis elegansLamin Gene in Nuclear Organization, Cell Cycle Progression, and Spatial Organization of Nuclear Pore Complexes

“…Coiled-coil interactions and head-to-tail associations between lamin monomers form 10-to 200-nm thick lamin filaments (reviewed in Stuurman et al, 1998) In vivo, lamin filaments are closely associated with the chromatin fibers (Belmont et al, 1993). In vitro, lamins can bind interphase chromatin (Hoger et al, 1991;Yuan et al, 1991;Taniura et al, 1995;Ulitzur et al, 1997;Goldberg et al, 1999a), mitotic chromosomes (Glass and Gerace, 1990;Glass et al, 1993), or specific DNA sequences (Shoeman and Traub, 1990;Luderus et al, 1992;Luderus et al, 1994;Baricheva et al, 1996;Zhao et al, 1996). The binding site of vertebrate lamins to chromatin is localized to specific sequences in the tail domain and can be displaced with the core histones H2A and H2B (Taniura et al, 1995;Goldberg et al, 1999a).…”

“…In vitro, lamins can bind interphase chromatin (Hoger et al, 1991;Yuan et al, 1991;Taniura et al, 1995;Ulitzur et al, 1997;Goldberg et al, 1999a), mitotic chromosomes (Glass and Gerace, 1990;Glass et al, 1993), or specific DNA sequences (Shoeman and Traub, 1990;Luderus et al, 1992;Luderus et al, 1994;Baricheva et al, 1996;Zhao et al, 1996). The binding site of vertebrate lamins to chromatin is localized to specific sequences in the tail domain and can be displaced with the core histones H2A and H2B (Taniura et al, 1995;Goldberg et al, 1999a).The composition of the nuclear lamina varies in different cell types and is under developmental regulation in both vertebrate and Drosophila cells (Stuurman et al, 1998). Metazoan cells contain between one to seven lamin proteins.…”