Nuclear envelope and chromatin compositional differences comparing undifferentiated and retinoic acid- and phorbol ester-treated HL-60 cells.
The human leukemic cell line (HL-60) can be induced to differentiate in vitro to granulocytic form with retinoic acid (RA), or to monocytic/macrophage form with phorbol ester (TPA). The granulocytic form acquires nuclear lobulation, nuclear envelope-limited chromatin sheets (ELCS), and cytoskeletal polarization, none of which are acquired following treatment with TPA. Immunoblotting analyses and capillary zone electrophoresis demonstrated that following RA treatment: lamins A/C and B1, and vimentin decreased to negligible amounts; LAP2 beta, lamin B2 and emerin remained essentially unchanged; lamin B receptor (LBR) increased markedly; histone subtypes H1.4 and 1.5 exhibited dephosphorylation. Following TPA treatment: lamins A/C and B1, B2 and vimentin increased in amount; LAP2 beta and emerin remained essentially unchanged; LBR increased markedly; histone subtypes H1.4 and 1.5 exhibited dephosphorylation. Emerin, which was cytoplasmic in undifferentiated or granulocytic cells, localized into the nuclear envelope following TPA. Normal human granulocytes revealed compositional differences compared to granulocytic forms of HL-60, namely increased vimentin and appearance of histone subtype H1.3. A working hypothesis for nuclear lobulation postulates a combination of: increased nuclear envelope deformability due to lamins A/C and B1 deficiency; an increase in nuclear surface area/volume; an increase in chromatin-nuclear envelope interactions.
Olins
A L
AL
Foundation for Blood Research, P.O. Box 190, 69 U.S. Route One, Scarborough, Maine 04070-0190, USA.
Herrmann
H
H
Lichter
P
P
Kratzmeier
M
M
Doenecke
D
D
Olins
D E
DE
D002451:Cell Compartmentation; D002454:Cell Differentiation; D002843:Chromatin / Q000737:chemistry*; D006098:Granulocytes / Q000737:chemistry; D018922:HL-60 Cells / Q000166:cytology / Q000187:drug effects; D006412:Hematopoietic Stem Cells / Q000737:chemistry / Q000166:cytology* / Q000187:drug effects; D006657:Histones / Q000032:analysis; D006801:Humans; D009685:Nuclear Envelope / Q000737:chemistry* / Q000648:ultrastructure; D009687:Nuclear Proteins / Q000302:isolation & purification; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014212:Tretinoin / Q000494:pharmacology*
D002843:Chromatin; D006657:Histones; D009687:Nuclear Proteins; D014212:Tretinoin; D013755:Tetradecanoylphorbol Acetate