Cellular protein kinase C isoform zeta regulates human parainfluenza virus type 3 replication.

Phosphorylation of the P proteins of nonsegmented negative-strand RNA viruses is critical for their function as transactivators of the viral RNA polymerases. Using unphosphorylated P protein of human parainfluenza virus type 3 (HPIV3) expressed in Escherichia coli, we have shown that the cellular protein kinase that phosphorylates P in vitro is biochemically and immunologically indistinguishable from cellular protein kinase C isoform zeta (PKC-zeta). Further, PKC-zeta is specifically packaged within the progeny HPIV3 virions and remains tightly associated with the ribonucleoprotein complex. The P protein seems also to be phosphorylated intracellularly by PKC-zeta, as shown by the similar protease digestion pattern of the in vitro and in vivo phosphorylated P proteins. The growth of HPIV3 in CV-1 cells is completely abrogated when a PKC-zeta-specific inhibitor pseudosubstrate peptide was delivered into cells. These data indicate that PKC-zeta plays an important role in HPIV3 gene expression by phosphorylating P protein, thus providing an opportunity to develop antiviral agents against an important human pathogen.

Mutant mice lacking the gamma isoform of protein kinase C show decreased behavioral actions of ethanol and altered function of gamma-aminobutyrate type A receptors.

Calcium/phospholipid-dependent protein kinase (protein kinase C, PKC) has been suggested to play a role in the sensitivity of gamma-aminobutyrate type A (GABAA) receptors to ethanol. We tested a line of null mutant mice that lacks the gamma isoform of PKC (PKC gamma) to determine the role of this brain-specific isoenzyme in ethanol sensitivity. We found that the mutation reduced the amount of PKC gamma immunoreactivity in cerebellum to undetectable levels without altering the levels of the alpha, beta I, or beta II isoforms of PKC. The mutant mice display reduced sensitivity to the effects of ethanol on loss of righting reflex and hypothermia but show normal responses to flunitrazepam or pentobarbital. Likewise, GABAA receptor function of isolated brain membranes showed that the mutation abolished the action of ethanol but did not alter actions of flunitrazepam or pentobarbital. These studies show the unique interactions of ethanol with GABAA receptors and suggest protein kinase isoenzymes as possible determinants of genetic differences in response to ethanol.

Specifications for bituminous road-mix surface dense-graded aggregate type, subclass C-1 : effective June 1, 1945.

Reproduced from typewritten copy.

Specifications for bituminous road-mix surface open-graded aggregate type, subclass C-2 (Bituminous retread-asphalt and tar) : effective May 15, 1939 ...

Cover-title.

Specifications for bituminous surface, open-graded aggregate type, subclass C-5 (Machine mix-emulsified asphalt) : effective July 1, 1945.

Reproduced from typewritten copy.

Specifications for bituminous surface dense-graded aggregate type, subclass C-6 (Machine mix) : effective September 1, 1937.

At head of title: ... U.S. Bureau of Public Roads' class C.

Specifications for bituminous surface, open-graded aggregate type, subclass C-4 (Machine mix-asphalt and tar) : effective May 15, 1939 ...

Cover-title.

The author's printing and publishing assistant, comprising explanations of the process of printing, preparation and calculation of manuscripts, choice of paper, type, binding, illustrations, publishing, advertising, & c. With an exemplification and description of the typographical marks used in the correction of the press.

Mode of access: Internet.

Organizational, direct support and general support maintenance repair parts and special tools lists : tamper, backfill, gasoline engine driven, hand-operated ram type (commercial construction equipment) model VR 11C, NSN 3895-01-151-2749 ; Stone Construction Equipment, Inc., contract number: DAA E07-82-C-6732.

Includes index.

Operator, organizational, direct support, and general support maintenance manual for air conditioner, vertical compact : type I, vertical, size C, 18,000 BTU/hr, class I, 208 volt, 3 phase, 50/60 hertz, Keco model F18T-2 : NSN 4120-00-168-1781.

"21 December 1979."

Operator, organizational, direct support and general support maintenance manual : heater, space, multifuel, with blower, 15,000 BTU/hr, 26 VDC, (Hunter model UH-48-C. type IV), NSN 4520-01-010-4059.

"August 1976."

Direct support and general support maintenance manual : water purification unit, van type, body mounted, electric motor driven, AC, DC, 115, 208 V, single and 3 phase, 60 hertz, 1/20 to 2 hp, 1500 gph, model EMC-1500S, NSN 4610-01-037-8746, EMC Industries Inc., DAAKO1-76-C-5661.

"1 August 1979."

Hodgkin's lymphoma cell lines express a fusion protein encoded by intergenically spliced mRNA for the multilectin receptor DEC-205 (CD205) and a novel C-type lectin receptor DCL-1

Classic Hodgkin's lymphoma (HL) tissue contains a small population of morphologically distinct malignant cells called Hodgkin and Reed-Sternberg (HRS) cells, associated with the development of HL. Using 3'-rapid amplification of cDNA ends ( RACE) we identified an alternative mRNA for the DEC-205 multilectin receptor in the HRS cell line L428. Sequence analysis revealed that the mRNA encodes a fusion protein between DEC-205 and a novel C-type lectin DCL-1. Although the 7.5-kb DEC-205 and 4.2-kb DCL-1 mRNA were expressed independently in myeloid and B lymphoid cell lines, the DEC-205/DCL-1 fusion mRNA (9.5 kb) predominated in the HRS cell lines ( L428, KM-H2, and HDLM-2). The DEC-205 and DCL-1 genes comprising 35 and 6 exons, respectively, are juxtaposed on chromosome band 2q24 and separated by only 5.4 kb. We determined the DCL-1 transcription initiation site within the intervening sequence by 5'-RACE, confirming that DCL-1 is an independent gene. Two DEC-205/DCL-1 fusion mRNA variants may result from cotranscription of DEC-205 and DCL-1, followed by splicing DEC-205 exon 35 or 34-35 along with DCL-1 exon 1. The resulting reading frames encode the DEC-205 ectodomain plus the DCL-1 ectodomain, the transmembrane, and the cytoplasmic domain. Using DCL-1 cytoplasmic domain-specific polyclonal and DEC-205 monoclonal antibodies for immunoprecipitation/Western blot analysis, we showed that the fusion mRNA is translated into a DEC-205/DCL-1 fusion protein, expressed in the HRS cell lines. These results imply an unusual transcriptional control mechanism in HRS cells, which cotranscribe an mRNA containing DEC-205 and DCL-1 prior to generating the intergenically spliced mRNA to produce a DEC-205/DCL-1 fusion protein.