Cell Recognition and Synapse Formation

Nirenberg, M.

Abstract

The nucleotide sequences of cDNA and genomic DNA clones for NK-1, NK-2, NK- 3 and NK-4 homeobox genes from Drosophila melanogaster were determined. NK-1 is expressed in small subsets of neurons and striated muscle cells in each thoracic and abdominal segment and in cells in the head. Genomic DNA was cloned from monkey and rat that corresponds to the Drosophila NK-1 homeobox DNA. Nucleotide sequence analysis showed that the amino acid sequences of the homeobox domains of monkey and rat are identical to that of the Drosophila NK-1 homeobox. NK-2 mRNA expression begins at the cellular blastoderm stage in the ventrolateral portion of the embryo as these cells become committed to the neuroectodermal pathway of development, which is the first step in the developmental program that leads to the formation of the nervous system. In 12 hr and older embryos NK-2 gene expression also was found in cells associated with the midgut. NK-3 and NK-4 are neighboring homeobox genes. NK-3 is expressed transiently in visceral mesoderm in 6-12 hr embryos in a segmentally repeated pattern. NK-4 mRNA also is expressed transiently starting as mesodermal cells appear during gastrulation (3 hr) and disappearing after 7.5 hr. Genomic DNA clones corresponding to 7 novel mouse homeobox genes were obtained and the nucleotide sequences of the homeobox regions were determined. Additional nucleotide sequence information was obtained for 2 of the 7 clones. Two novel species of Pou box-homeobox cDNA were cloned and the Pou box-homeobox regions were sequenced. Clones of mouse genomic DNA fragments with enhancer activity in N18TG2 mouse neuroblastoma cells were obtained by a selection method. CTF/NF-1 and LVc proteins extracted from N18TG2 nuclei bind to sites in cloned N1 DNA and function as enhancers. cDNA clones were obtained that correspond to species of NG108-15 poly A + RNA that increase in abundance when cells are treated with compounds that elevate cells cAMP. Four of the clones correspond to different regions of mitochondrial DNA, one clone corresponds to secretogranin I, and one corresponds to ribosomal protein S-10 mRNA.

Funding Agency

• Agency: National Institute of Health (NIH)
• Institute: National Heart, Lung, and Blood Institute (NHLBI)
• Type: Intramural Research (Z01)
• Project #: 1Z01HL000009-16
• Application #: 3878867
• Support Year: 16
• Fiscal Year: 1990

Institution

• Name: National Heart, Lung, and Blood Institute
• Country: United States

Publications

Xia, Menghang; Huang, Ruili; Guo, Vicky et al. (2009) Identification of compounds that potentiate CREB signaling as possible enhancers of long-term memory. Proc Natl Acad Sci U S A 106:2412-7

Sharma, Shail K; Nirenberg, Marshall (2007) Silencing of genes in cultured Drosophila neurons by RNA interference. Proc Natl Acad Sci U S A 104:12925-30

Koizumi, Keita; Higashida, Haruhiro; Yoo, Siuk et al. (2007) RNA interference screen to identify genes required for Drosophila embryonic nervous system development. Proc Natl Acad Sci U S A 104:5626-31

Chen, Xiaobing; Vinade, Lucia; Leapman, Richard D et al. (2005) Mass of the postsynaptic density and enumeration of three key molecules. Proc Natl Acad Sci U S A 102:11551-6

Ivanov, Andrej I; Rovescalli, Alessandra C; Pozzi, Paola et al. (2004) Genes required for Drosophila nervous system development identified by RNA interference. Proc Natl Acad Sci U S A 101:16216-21

Lum, Lawrence; Yao, Shenqin; Mozer, Brian et al. (2003) Identification of Hedgehog pathway components by RNAi in Drosophila cultured cells. Science 299:2039-45

Nirenberg, Marshall (2003) The genetic revolution: the importance of flies and worms. Am J Psychiatry 160:615

Koizumi, Keita; Lintas, Carla; Nirenberg, Marshall et al. (2003) Mutations that affect the ability of the vnd/NK-2 homeoprotein to regulate gene expression: transgenic alterations and tertiary structure. Proc Natl Acad Sci U S A 100:3119-24

Hwang, Kae-Jung; Xiang, Bosong; Gruschus, James M et al. (2003) Distortion of the three-dimensional structure of the vnd/NK-2 homeodomain bound to DNA induced by an embryonically lethal A35T point mutation. Biochemistry 42:12522-31

Wang, Lan-Hsiang; Chmelik, Rebecca; Nirenberg, Marshall (2002) Sequence-specific DNA binding by the vnd/NK-2 homeodomain of Drosophila. Proc Natl Acad Sci U S A 99:12721-6