RNA synthesis in cells infected with herpes simple virus. XIII. Differences in the methylation patterns of viral RNA during the reproductive cycle

We compared the rates of synthesis of herpes simplex virus type 1 glycoproteins C and D and quantitated the accumulation of translatable mRNA for each glycoprotein at various times after infection. The rate of synthesis of gD increased sharply early in the infection, peaked by 4 to 6 h after infection, and declined late in the infection. In contrast, the rate of synthesis of gC increased steadily until at least 15 h after infection. The levels of mRNA for both of these glycoproteins, as detected by hybridization and by translation in vitro, continued to increase until at least 15 or 16 h after infection. Synthesis of both gC and gD and their respective mRNAs was found to be sensitive to inhibition of viral DNA replication with phosphonoacetic acid. The finding that reduced amounts of gD were synthesized late in the replicative cycle, whereas gD mRNA continued to accumulate in the cytoplasm, argues that the synthesis of gD is regulated, in part, at the level of translation.

Section: Discussionmentioning

confidence: 99%

Evidence for translational regulation of herpes simplex virus type 1 gD expression

Johnson¹,

Spear²

1984

“…HSV-1 mRNA shares general properties with host cell mRNA; i.e., it is synthesized in the nucleus, polyadenylated on the 3' end, capped on the 5' end, and internally methylated (7,9,37,46,49,58). It is, therefore, interesting to note that HSV-1 mRNA appears to differ from most mammalian mRNA in its degree of splicing, especially in the interior of mRNA coding regions.…”

Section: Discussionmentioning

confidence: 99%

Herpes simplex virus type 1 HindIII fragment L encodes spliced and complementary mRNA species

Frink¹,

Anderson²,

Wagner³

1981

We have used DNA bound to cellulose to isolate and translate in vitro herpes simplex virus type 1 (HSV-1) mRNA's encoded by HindIII fragment L (mapping between 0.592 and 0.647), an 8,450-base-pair (8.45-kb) portion of the long unique region of the viral genome. Readily detectable, late mRNA's 2.7 and 1.9 kb in size encoding 69,000-and 58,000-dalton polypeptides, respectively, were isolated. A very minor late mRNA family composed of two colinear forms, one 2.6 kb and one 2.8 kb, was isolated and found to encode only an 85,000-dalton polypeptide. A major early mRNA, 1.8 kb in size encoding a 64,000-dalton polypeptide, was also isolated. High-resolution mapping of these mRNA's by using Si nuclease and exonuclease VII digestion of hybrids between them and 5' and 3' end-labeled DNA fragments from the region indicated that the major early mRNA contained no detectable splices, and about half of its 3' end was complementary to the 3' region of the very minor 2.6-to 2.8-kb mRNA's encoded on the opposite strand. These mRNA's also contained no detectable splices. The major late 2.7-kb mRNA was found to be a family made up of members with no detectable splices and members with variable-length (100 to 300 bases) segments spliced out very near (ca. 50 to 100 bases) the 5' end. Like all herpesviruses, herpes simplex virus type 1 (HSV-1) is characterized by an unusual arrangement of its genome. The viral DNA has a molecular weight of 95 x 106 to 100 x 106 daltons (d; reviewed in reference 44), which corresponds to a length of 150,000 base pairs (150 kb; 50). The linear HSV-1 genome is segmented into a long unique region (UL, ca. 105 kb, 70%)

“…mRNA molecules of HSV-1 associated with the polyribosomes of infected cells resemble mRNA molecules of other DNA viruses and eucaryotic cells. HSV-1 mRNA, in addition to being polyadenylated, is capped at the 5' end (4,20), and we have found its average size to be no less than 2 kb (28). However, due in part to the large size and complexity of the HSV genome, little has been reported concerning the properties and origins of individual viral mRNA species.…”

mentioning

confidence: 89%

Isolation and localization of herpes simplex virus type 1 mRNA

Anderson¹,

Stringer²,

Holland³

et al. 1979

Herpes simplex virus (HSV) DNA bound to cellulose has been used as a reagent to isolate viral mRNA for size analysis on denaturing agarose gels. Total viral polysomal polyadenylated RNA was isolated from cells late after infection when such RNA has sequences encoded by approximately 45% of the HSV DNA. This RNA has a size range of from 1.5 to-8 kilobases, with certain sizes, such as 1.7 to 1.9 kilobases, being favored. We have used the restriction endonucleases HindIII and XbaI singly and together to generate various sized fragments covering the entire HSV-1 genome. These fragments have been bound to cellulose to allow isolation of HSV-1 mRNA annealing to different regions of the viral genome. Discrete sizes of viral mRNA are associated with certain regions of the genome, but the mRNA population hybridizing to even the smallest restriction fragments is complex. We used hybridization of size-fractionated RNA to Southern blots of restriction fragments of HSV-1 DNA generated by the BglII as well as HindIII and XbaI endonucleases to confirm the preparative hybridization data and to provide some overlap data for positioning transcripts. The data of blot and preparative hybridization agreed very well and were combined to construct a preliminary transcription map of HSV-1. Such a map revealed at least two areas of the long unique region of the HSV-1 genome which annealed to a large number of HSV-1 transcripts. Furthermore, discrete-sized mRNA species larger than 5 kilobases in length were found only in the middle of the long unique region. The implications of these data are discussed.